HLATools™ - To Parent Directory
On-Line Manual
HLATools™
HLATools™ v. 1.1
June 2006
For Research Use Only. Not for Use in Diagnostic Procedures.
20001 Kittridge Street, Canoga Park, CA 91303-2801
Tel: (818) 702-0042 Fax: (818) 702-6904
www.onelambda.com
Advancing Transplant Diagnostics
Manual_Template_R1
OLI-UD-0156-062006
HLATools_RUOManual_v1.1_Rev. 0.pdf
©Copyright
2006 One Lambda, Inc.
LABType is a registered trademark of One Lambda, Inc.
™HLATools is a trademark of One Lambda, Inc.
™LABScan 100 is a trademark of One Lambda, Inc.
®
Luminex is a registered trademark of Luminex Corporation.
®Windows is a registered trademark of Microsoft Corporation.
®
All One Lambda software products are designed to assist personnel
experienced in HLA analysis by suggesting typing results. However, any
clinical or diagnostic results must be carefully reviewed by a person qualified
in HLA typing to assure correctness. The software may be used to aid in
suggesting results, but should not be used as the sole method for determining
reportable results. The software is meant as a laboratory aid, not as a source
of definitive results. The software design does not mitigate hazards
associated with the software. The laboratory director or technologist trained in
histocompatibility testing is required to review all data to detect any problems
with the software.
Table of Contents
Chapter 1:
Introduction to One Lambda HLATools™
Overview. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
HLATools Modules . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
HLATools Configuration Suite . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
HLA Update . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Comments Editor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Configuration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Connections Tool . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Database Tools . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
System Diagnostics. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
User Management . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
About this Manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
How to Use This Guide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
Viewing the HTML Version of this Manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Conventions Used in this Manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Interface Nomenclature. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Technical Support . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Chapter 2:
Installation and Log On
Software Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
System Requirements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
Luminex Software Compatibility . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Software Installation on all User Computers . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Creating the LuminexFiles Folder . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Running the Clean Database Utility . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Modifying, Repairing or Uninstalling HLATools . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
Modifying HLATools . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
Repairing HLATools . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
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Uninstalling HLATools . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
Complete Uninstall of HLATools. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
Updating HLATools . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Log On . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
Chapter 3:
LABType Home
Product Inserts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Patient Worksheets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Bead Probe Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Resolution Limitations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
Chapter 4:
Lambda Explorer
Accessing Luminex .csv Files . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
Importing Luminex .csv Files . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
Chapter 5:
HLATools™ LABType Interactive
Synopsis of LTI Batch Analysis Views . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Batch Data Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
Batch Results View. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
Bead Analysis View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
Control Values View – Positive Controls. . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
Control Values View – Minimum Bead Count . . . . . . . . . . . . . . . . . . . . . . . 45
Synopsis of Sample Analysis Views and Subviews . . . . . . . . . . . . . . . . . . . . . . . . . . 46
Sample Results Analysis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Assignments View with Matched Reaction Pairs and Close Reactions . . . . . . . . 47
Matched Reaction Pairs Subview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
All Alleles Subview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
Type/Subtype Subview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
Closest Reactions Subview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
Specificity Subview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Sample Assignment Subview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Patient Information Subview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
Bead Analysis (Sample) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
Reaction Grid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
Sorting the Reaction Grid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
Clustering Positives to the Left of the Grid . . . . . . . . . . . . . . . . . . . . . . . . . . 61
Restoring Allele Order . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
Shifting Alleles to the Top of the Grid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
Restoring Allele Order . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
Sorting by Bead Usage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
Sorting by Allele Pattern Matches . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
Raw Data Table . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
Chapter 6:
Patient Explorer
Overview. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
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Chapter 7:
Reference Files and Their Maintenance
Survey of LTI Data Resource Files . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
Products > View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
Products > Update. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
NMDP > View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
NMDP > Update . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78
Serological > View . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78
Serological > Update . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
Filter > View. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
Chapter 8:
HLATools™ Lambda Reporter
Laboratory Report Header Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82
Entering Laboratory Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83
The other fields are self-explanatory and do not require elaboration. . . . . . . . . . 84
Using Lambda Reporter Filters. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
Selecting Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
Creating a Custom Report Template . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88
Report Sections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90
Specifying Report Paper Size . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
Chapter 9:
HLATools™ Legacy Reporter
Preformatted Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93
Patient Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
1B – Patient Summary Report Filtered by Batch . . . . . . . . . . . . . . . . . . . . . . 94
2A – Patient Report. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94
Batch Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
3A – Batch Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95
3B – Batch Raw Data Abbreviated Specificity Report. . . . . . . . . . . . . . . . . . 96
3D – Batch Report Sorted by Patient . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96
3E – Batch Report Sorted by Sample . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97
3F – Combined Batch Report Sorted by Sample . . . . . . . . . . . . . . . . . . . . . . 97
3G – Combined Batch Report Sorted by Patient . . . . . . . . . . . . . . . . . . . . . . 98
Sample Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98
3C – Sample Report Filtered by Batch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98
4A – Sample Raw Data Abbreviated Specificity Filtered by Batch. . . . . . . . 98
4B – Combined Sample Analysis Data Report. . . . . . . . . . . . . . . . . . . . . . . . 99
4C – Sample Raw Data Abbreviated Specificity Report . . . . . . . . . . . . . . . 100
4D – Sample Abbreviated Specificity Report. . . . . . . . . . . . . . . . . . . . . . . . 100
4E – Sample Complete Specificity Report . . . . . . . . . . . . . . . . . . . . . . . . . . 100
99A – Sample Report with Chart Filtered by Batch . . . . . . . . . . . . . . . . . . . 100
Summary Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101
1A – ABDR by Batch Sorted by Well Position . . . . . . . . . . . . . . . . . . . . . . 101
1C – Batch Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101
5B – Allele Query Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101
6A – Serological Equivalent Table Report . . . . . . . . . . . . . . . . . . . . . . . . . . 102
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6B – NMDP Allele Code Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102
6C – Batch Data File Summary Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102
6E – Catalog Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102
Special Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103
5A – Allele Group Frequencies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103
6D – Database Table Information. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103
99B – ABDR by Batch Sorted by Well Position . . . . . . . . . . . . . . . . . . . . . 103
Appendix A: LTI Interface Customization
Modifying Tables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107
Changing Sort Order or Grouping . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107
Changing Batch Results Column Order . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108
Navigating the LTI Interface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109
Navigating Between Windows Using the Keyboard . . . . . . . . . . . . . . . . . . 109
Accessing LTI Functions via Hot Keys . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109
Changing Date Formats in HLATools . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109
Editing the Assignment Subview Comments List . . . . . . . . . . . . . . . . . . . . . . . . . . 109
Specifying the Data Transfer File Export Type . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110
Specifying the Character Encoding Type . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
Appendix B: HLATools™ FAQ
Frequently Asked Questions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113
How can I tell if HLATools is using the most recent LABType products? . . . . 113
Why can’t I find the Admin Diagnostics tool? . . . . . . . . . . . . . . . . . . . . . . . . . . 113
Update Database – Why can’t I run …? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114
LTI database does not exist – Why do I get a message that …? . . . . . . . . . . . . 114
Remote LTI database – Why can’t I connect to … using Windows XP? . . . . . 114
Open a port though the XP firewall – How do I …? . . . . . . . . . . . . . . . . . . . . . 114
Local security policy on XP – How do I modify … . . . . . . . . . . . . . . . . . . . . . . 116
Clean Database – Why do I get a BCP error …? . . . . . . . . . . . . . . . . . . . . . . . . 116
Why do I have to log into the database when I try to print some reports? . . . . . 118
Windows Authentication Mode – Why can’t I set up …? . . . . . . . . . . . . . . . . . 118
Alleles – How are they designated? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 118
Matched Reaction Pairs (MRPs) – How does LTI identify …? . . . . . . . . . . . . . 120
MRP Table – How does LTI populate the …? . . . . . . . . . . . . . . . . . . . . . . . . . . 122
MRP Table – How are Allele1 and Allele2 grouped in the …? . . . . . . . . . . . . . 124
All Alleles Table – How is the … generated? . . . . . . . . . . . . . . . . . . . . . . . . . . 126
Type/Subtype Allele Assignments – How are … made? . . . . . . . . . . . . . . . . . . 127
Reaction Grid – How is the … sorted? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128
NMDP Codes – How can I automatically update …? . . . . . . . . . . . . . . . . . . . . 130
LTI database directory – Why is the … growing excessively? . . . . . . . . . . . . . 131
How do I update an LTI database with a non-default name? . . . . . . . . . . . . . . . 132
How do I install MSDE to a non-default folder? . . . . . . . . . . . . . . . . . . . . . . . . 133
How can I import LABType Classic analyses into HLATools? . . . . . . . . . . . . 134
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Appendix C: Network Installation of HLATools
Peer-to-Peer Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137
Creating a Database on a Remote Server . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 138
Connecting to a Remote LTI Database. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 140
Editing the Authorized Users List from a Remote Server . . . . . . . . . . . . . . . . . 148
Linking to another Server using SQL Authentication . . . . . . . . . . . . . . . . . . . . 149
Setting up HLATools in a Client/Server Network (to come) . . . . . . . . . . . . . . . . . . 151
HLATools Security Modes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 151
Sharing Luminex Files on a Network . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152
Glossary
Glossary Entries . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157
A260/A280 Ratio . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157
ACD . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157
Allele Name Extensions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157
Amino Acid Codes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157
ASO . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158
Class I MHC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158
Class II MHC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158
Cell death detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159
Clean Database . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160
Close Reaction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160
Code definition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160
Complement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160
Count Type . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160
CREG . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160
EDTA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
ELISA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
Epitope . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
False Negative. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
False Positive . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
False Reaction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
Genetic Code. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 162
Immunomagnetic beads . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 162
JPN Rank . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 162
Local Code . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 162
Log Files [.ldf] . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 162
Matched Reaction Pair . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
Mean and Median . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
Merge Database . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
MIC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
N80/K80 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
Negative Control. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
NIH Score . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 164
NMDP code . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 164
Normalization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 164
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Nucleotide Codes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165
Null Allele. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165
PBS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165
PCR-SSP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165
Positive Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 166
Positive Reaction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 166
Primary Data Files [.mdf] . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 166
Recognition Site . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 166
RFLP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
Secondary antibody. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
SSO . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
Substrate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
Unicode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
Update Database . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
Index
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Chapter 1
Introduction to One Lambda
HLATools™
This manual is intended to serve as in-depth discussion of the features of the One Lambda HLATools™ software suite.
This manual supersedes the introductory topics that were treated in Getting Started with One Lambda HLATools
and contains advanced topics and reference material that goes beyond the scope of that document.
All One Lambda software products are designed to assist personnel experienced in HLA analysis by suggesting typing
results. However, any clinical or diagnostic results must be carefully reviewed by a person qualified in HLA typing to
assure correctness. The software may be used to aid in suggesting results, but should not be used as the sole method for
determining reportable results. The software is meant as a laboratory aid, not as a source of definitive results.
Overview
One Lambda HLATools is a suite of software applications that interpret the results obtained by One Lambda’s LabType
SSO HLA typing tests. The software suite contains five major modules and a number of utilities.
HLATools Modules
The application suite comprises the following modules:
LABType Home – a repository of One Lambda product/lot-specific information in PDF format for LABType SSO and
Micro SSP product lines.
Lambda Explorer – a utility for importing files produced by the Luminex LABScan<SuperScript>™ 100 bench-top
flow analyzer into the LabType Interactive (LTI) database.
LabType Interactive – a software application that contains tools for assessing the quality of imported data, reviewing
preliminary analysis results, adjusting cutoff values and accepting suggested allele assignments.
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Patient Explorer – a utility that collates multiple sample results collected from among one or more batches with a
single patient identified by a unique patient ID.
Lambda Reporter – a report generator that creates a variety of preformatted batch, sample and patient reports. The
Lambda Reporter also exports special reports such as ABDR reports as well as files in .xls, .txt, .pdf and .doc formats.
HLATools Configuration Suite
The HLATools Configuration Suite interface provides access to a number of utility programs used to configure the
HLATools modules. To access these programs, select Start > Programs > One Lambda > HLATools Configuration
Suite.
HLA Update
•
Products modifies how LTI handles data on a per-product basis:
•
Specifies type of mean and type of bead count used in data handling
•
Specifies minimum bead count and positive control value thresholds
•
Establishes order in which products appear in pick list when user matches a product to session data
•
Specifies cutoff sensitivity used to classify strength of bead readings
For details, see Products > View, p. 72.
•
Update LABType product information from files downloaded from the One Lambda website
For details, see Products > Update, p. 75.
•
NMDP maintains NMDP ambiguity code listing
•
Displays code list
For details, see NMDP > View, p. 77.
•
Updates code list from NMDP Research website or a local file
For details, see NMDP > Update, p. 78.
•
Serological maintains allele/serological equivalents list
•
Displays equivalents list
•
Exports list in plain .csv or in Excel-compatible .csv format
For details, see Serological > View, p. 78.
•
Updates equivalents list from a website or a local file
Comments Editor
•
Constructs list of frequently used comments for user-supplied sample annotation
For details, see Figure A-3, HLATools Comments Editor Tool.
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Configuration
•
General Configuration
•
Lambda Explorer Default Directory specifies the pathway to the default folder containing Luminex 100 output files accessed by the Lambda Explorer (see Chapter 4).
•
Lab Info maintains laboratory and institutional information included in report headers
For details, see Entering Laboratory Information, p. 83.
•
•
•
Login Type sets level of HLATools login security (see HLATools Security Modes, p. 149).
LABType
•
Export Type specifies client-specific data transfer file export types and enables client-specific application features.
•
Character Encoding Type specifies the character encoding (code page) used in report generation.
LABScreen
•
Specifies the pathway to the folder containing the One Lambda LABScreen HLA analysis executable. LABScreen can be launched from HLATools explorer bar (Figure 5-1).
Connections Tool
•
Provides access to multiple LTI databases on networked servers.
For details, see Connecting to a Remote LTI Database, p. 138.
Database Tools
•
Attach/Detach makes or breaks connections to LTI databases. For details about the need to detach databases, see
the FAQ on excessive growth in database size, Appendix B, p. 129.
•
Backup/Restore displays the current instance of the SQL server and the attached LTI database and backs up an existing LTI database or restores a backed-up database located on a local or a remote server.
•
Clean/Update creates a “clean” or new LTI database or updates an existing one. See the Clean Database and Update Database Glossary topics.
•
Merge combines two existing LTI databases. See the Merge Database Glossary topic.
System Diagnostics
•
Displays information about
•
current user and users authorized to use local computer
•
local computer OS, hardware components and IP address
•
creation dates and last time of use of program components
•
system environment variables
•
currently implemented LTI database connections
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User Management
•
Creates new users authorized to access a local or remote SQL database
•
Adds Windows User and Groups authorized to access databases
For details, see Figure C-2, User Management Tool.
Features
The HLATools software suite is designed to be used with One Lambda’s LabType SSO HLA typing tests. The central
module in the suite is the LabType Interactive (LTI) analysis tool. The bulk of this document is devoted to explaining
the use and features of LTI. With LTI and the other software modules in this suite, you can accomplish the tasks listed
below.
•
Import data from the LABScan™ 100 bench-top flow analyzer (Importing Luminex .csv Files, p. 31).
•
Review preliminary results in tabular format (Batch Data Analysis, p. 36).
•
Assess the quality of the imported data by comparing it to demographic reference data (Bead Analysis View, p. 42).
•
Adjust cutoff values globally and locally, as warranted (Bead Analysis View, p. 42 and Assignments View with
Matched Reaction Pairs and Close Reactions, p. 47).
•
Accept suggested allele assignments (Assignments View with Matched Reaction Pairs and Close Reactions, p. 47).
•
Review identified Close Reactions, cases where reversing a single bead reaction (making a negative positive or
vice versa) would yield a Matched Reaction Pair and an allele assignment. (Closest Reactions Subview, p. 55)
•
Review types and subtypes of assigned alleles, as well as ambiguous cases where the beads cannot determine with
precision which specific alleles are present and allele assignments cannot be made. (Type/Subtype Subview, p. 54)
•
Review allele specificities for any given bead in the sample (Specificity Subview, p. 56)
•
Identify and assign serological equivalents to the assigned allele (All Alleles Subview, p. 53 and Sample Assignment
Subview, p. 56)
•
Analyze sample reaction patterns in a serogram-like grid (Reaction Grid, p. 58)
•
Review raw data in a tabular format (Raw Data Table, p. 64)
•
Manage data from multiple samples associated with a single patient (Chapter 6, Patient Explorer)
•
Update the data resource files used by LabType Interactive (Chapter 7, Reference Files and Their Maintenance)
•
Generate a wide variety of sample, patient and batch reports (Chapter 8, HLATools™ Lambda Reporter and
Chapter 9, HLATools™ Legacy Reporter)
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About this Manual
This manual is intended to provide the basic information a user needs to take advantage of the advanced features of
LABType Interactive and the other modules in the HLATools software suite, and to understand the purpose of each
element contained in the module interfaces.
It is assumed that the reader has read and understood one of One Lambda’s LABType SSO Product Inserts and is
familiar with the basics of PCR amplification and data collection using the Luminex LABScan<SuperScript>™ 100
Flow Analyzer. A number of terms pertinent to LTI and other HLATools modules, and to other One Lambda products
are contained in the Glossary for easy reference. Details about laboratory aspects of HLA typing, LabType assays or
statistical analysis are beyond the scope of this manual.
How to Use This Guide
This guide is divided into three principal parts: these introductory remarks, software installation and networking
instructions, and usage instructions for the various HLATools software modules. The manual also includes appendices
dealing with topics in interface customization, FAQs, multi-user installations, a Glossary, and an Index.
Page numbers in the on-line version refer to page numbers in the printed version of the guide.
Chapter 2, Installation and Log On, contains step-by-step instructions for installing the software on a single-user
standalone workstation.
Chapter 3, LABType Home, provides a brief survey of the One Lambda product literature that forms the basis of much
of the allele specific information that is contained in your HLATools database.
Chapter 4, Lambda Explorer, describes how to access Luminex files and import them into the HLATools database.
Chapter 5, HLATools™ LABType Interactive, describes how to use the LTI software application to analyze batch and
individual patient or sample results and how to interpret allele assignments.
Chapter 6, Patient Explorer, describes how to collate and review the results of multiple sample tests that have been
carried out on a single patient.
Chapter 7, Reference Files and Their Maintenance, describes the contents of several important data files which form
part of the HLATools database and how to update them.
Chapter 8, HLATools™ Lambda Reporter, provides an overview of the tools available to create user- and laboratoryspecific custom reports.
Chapter 9, HLATools™ Legacy Reporter, provides an overview of the different types of preformatted sample, patient,
batch, summary and special reports available to the HLATools user.
Appendix A, LTI Interface Customization, describes some techniques that users can employ to change the appearance
of charts and tables in LTI.
Appendix B, HLATools™ FAQ, addresses a number of Frequently Asked Questions.
Appendix C, Network Installation of HLATools, deals with establishing user access in different configurations such as
standalone workstations, peer-to-peer installations and multi-user network environments.These instructions deal with
some common scenarios that you may encounter when using LTI in a multi-user laboratory environment.
A small Glossary of terms pertinent to HLA typing and screening is included.
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The printed manual concludes with an Index.
This manual has been designed to be printed out or viewed online. The distribution media contains electronic versions
of this manual in PDF and HTML format.
Viewing the HTML Version of this Manual
When viewing the HTML version of this manual, the Internet Explorer browser may repeatedly generate a message
warning that the files contain active content. You can suppress this message by reconfiguring Internet Explorer as
follows:
1. Select Start > Settings > Control Panel > Internet Options > Advanced.
2. Deselect the Security option “Allow active content to run in files on My computer.”
You may wish to re-enable this option when you have finished with your HLATools session.
Conventions Used in this Manual
This manual assumes that you are familiar with the Windows operating system and with the standard desktop functions
that are common to virtually all software programs:
•
Basic file operations (opening, saving and printing)
•
Editing functions (cutting, copying and pasting text)
•
Resizing windows and dialog boxes
•
Simple manipulation of tables
It also assumed that you are familiar with the standard Windows file and folder tree structure and that you know how to
manipulate folders and the files they contain.
Menu options and dialog box selections in step-by-step procedures are indicated in green text with a right angle bracket
marking each step. A step may involve actions such as making a menu selection, accessing a pane through a tab, or
checking a check box or radio button. For example, to access the HLATools application, select Start > Programs >
One Lambda > HLATools > HLATools.
Names of Views, Subviews, Panels, Dialog Boxes, and Buttons are shown in boldface type except when they form
part of a hypertext cross-reference or a navigation path to a particular view, subview or panel. When used as hypertext
they appear in blue or green type: Conventions Used in this Manual.
Interface Nomenclature
•
The term View is used instead of “window” or “tab”.
•
A tabbed view is termed a View or Subview.
•
A Panel or a Pane is a demarcated region or area in a View or Subview.
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•
A Batch is a collection of Samples and appears as a folder in the Batch/Samples file tree. Note that the contents of
a batch are termed “samples” and not “patients”. This distinction is made because multiple samples may be associated with a single patient, and because the confidentiality policies of some institutions require that the patient/sample relationship not be openly divulged. The treatment of patients with multiple samples is addressed in Chapter 6,
Patient Explorer.
•
The term Bead is often used synonymously (but imprecisely) with Probe since a LABType bead can be assumed to
carry only one type of probe.
Technical Support
For technical support or to report software problems, contact your One Lambda representative.
•
From the United States, call 800-822-8824.
•
In the greater Los Angeles area, call 818-702-0042.
•
Contact us by e-mail at: [email protected]
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Chapter 2
Installation and Log On
This chapter addresses installation of HLATools™ software on a single-user standalone workstation. Installation of
HLATools software in a peer-to-peer configuration or client/server network environment is discussed in Appendix C,
Network Installation of HLATools.
Software Installation
System Requirements
These are system requirements for installation and use of HLATools software:
•
Microsoft® Windows® 2000 (SP4) or XP (SP2 or later) operating system
•
Microsoft .NET Framework v. 1.1
•
Microsoft SQL Server Desktop Engine 2000 (SP3a) – this must also be installed on client computers in a client/server configuration
•
Pentium® III with Windows 2000 or Pentium 4 with Windows XP
•
256 MB of RAM
•
5 GB of available hard disk space
•
CD-ROM drive
All software needed to install HLATools on standalone or client computers is contained on the CD-ROM in your
software shipment. The installation of the SQL database server software should be handled by your IT system administrator. The MS Windows SQL Server software must include Microsoft SQL Server 2000 Service Pack 3a which is
available from Microsoft at http://www.microsoft.com/downloads/.
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Luminex Software Compatibility
HLATools software has been developed and tested to be compatible with Luminex IS 2.1 and IS 2.2 output files
produced by the LABScan 100 flow analyzer.
Software Installation on all User Computers
1. You must have administrator status on the PC in order to install HLATools and the HLATools Configuration Suite.
2. Place the CD-ROM into your CD-ROM drive.
3. Many Windows computers are configured so that CDs auto-run by default. In the event that auto-run has been disabled on your system, select Start > Run > Browse to locate your CD-ROM drive.
4. Double-click on the CD-ROM icon to launch the installation program.
5. If the Program Maintenance dialog box displays the following message, you are reinstalling HLATools over an
existing installation. In that case, proceed to Modifying, Repairing or Uninstalling HLATools, p. 19; otherwise,
continue with the next step.
Figure 2-1: InstallShield Modify… Message
6. Once the installation begins, you may see this message:
Figure 2-2: Microsoft .NET Framework Installation
This occurs only if the software detects that Microsoft .NET Framework has not been installed on your computer.
If Microsoft .NET Framework is already installed, this step of the install is skipped.
If the installation software cannot detect the Microsoft .NET Framework software, it will launch the InstallShield
for Microsoft .NET Framework. Accept the license agreement, click Install and wait for the installation to be
completed.
7. Next, the installation software prepares for the actual installation of the HLATools software package. When the
preparation steps have been completed, you may be requested to reboot your computer. If so, click Restart and
continue. After Windows restarts, the next step in the installation may last several minutes.
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8. When the HLATools Setup InstallShield Wizard appears, click Next, read and accept the license agreement and
click Next again to continue.
Figure 2-3: Customer Information Dialog
9. Regarding the Customer Information dialog:
A. The User Name field is sometimes filled in automatically by your computer.
B. An entry in the Organization field is optional.
C. It is recommended that the default option that allows any user of the computer to use the HLATools software be
accepted.
Click Next to proceed.
10. Two Setup Type options are available.
A. Complete – installs all program features and requires about 260 MB. The default installation destination is
your C: drive.
B. Custom – allows users to select installation options:
•
Install HLATools to a destination other than the C: drive;
•
Check available drive space.
The default option is the complete installation. Click Next to proceed.
11. The InstallWizard is now ready to complete the installation.
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Click Install to proceed. If for some reason you change you mind, it is possible to abort the installation while it is
in progress. Click Finish to conclude the installation.
This completes the first part of HLATools software installation for a single workstation. Before running HLATools, you
must create a folder where your Luminex files can reside and run the Clean Database utility to create an HLATools
database structure. See Creating the LuminexFiles Folder, p. 18, and Running the Clean Database Utility, p. 18, for
further instructions.
Creating the LuminexFiles Folder
When first launched, the Lambda Explorer will look at the top level of the directory structure where HLATools has
been installed for folders containing files in the .csv format. This is the format used for the Luminex
LABScan<SuperScript>™ 100 output files that are the input files for the LTI analysis module. In a standard installation to the C: drive of your computer, the Lambda Explorer will look for a folder called LuminexFiles. As the first
step of the installation, create the folder for your .csv files as follows:
1. Select My Computer >Local Disk (C:).
2. From the desktop menu select File > New > Folder.
3. Name the folder exactly LuminexFiles and press Enter. Use this folder to store your Luminex LABScan 100 .csv
files.
If you have installed the HLATools software somewhere other than in your C: drive, you can use the built-in browse
function to locate .csv files for analysis.
For instructions on how to access Luminex output files directly from the computer that controls your Luminex
analyzer, see Sharing Luminex Files on a Network, p. 150.
Running the Clean Database Utility
This utility creates the database structure required by LTI and other HLA Tools applications. The database files are
located in the folder C:\Program Files\Microsoft SQL Server\MSSQL$ONELAMBDA.
Warning: If you are installing HLATools for the first time, you must run the Clean Database utility before launching
any of the HLATools applications. You must have administrative permissions in order to run Clean Database. If you
attempt to use the Lambda Explorer to import files or run LTI without previously creating a database, both applications
will generate a warning message and quit. If you are updating HLATools, you probably want to preserve your existing
analysis results. To determine if you should run Clean Database or Update Database, see Updating HLATools, p. 23.
To run Clean Database:
1. Select Start > Programs > One Lambda > HLATools Configuration Suite > Clean/Update.
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2. The Clean Database utility takes several minutes to create and populate the database structure. Its progress is displayed in a log message window.
Figure 2-4: Clean DataBase Log Message Window
Warning: Running Clean Database creates a new, clean database, which means that all previously analyzed samples
and patient information overwritten when the new database is created. All current patient/sample associations and
allele assignments will be lost.
Modifying, Repairing or Uninstalling HLATools
The HLATools InstallShield utilities provided on the distribution CD allow you to modify, repair or uninstall
HLATools. You must have administrative permissions in order to perform these functions. These utilities affect only
the HLATools program files and do not alter your Luminex data files or your database.
To use the program maintenance utilities:
1. Insert the HLATools CD and allow it to AutoStart. Alternatively, locate and launch the setup utility:
2. Click Next to proceed. The Program Maintenance dialog will appear (Figure 2-5). The InstallShield Wizard
allows you to modify, repair or remove One Lambda HLATools.
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Figure 2-5: Program Maintenance Dialog
Modifying HLATools
To modify HLATools software applications:
1. Select the Modify option shown in Figure 2-5:
2. Click Next to access the Custom Setup dialog (Figure 2-6)
•
Installation options that will be offered in future releases can be accessed via the pull-down menu.
•
If you select an option and the message in the Feature Description field declares that the feature requires no
space on your hard drive, this means that the feature is already installed.
3. Click Next to proceed, then Install, then Finish to carry out the modification.
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Figure 2-6: Custom Setup Options
Repairing HLATools
If you encounter a runtime error message similar to that shown in Figure 2-7, you can run the Repair utility without
completely reinstalling the program.
Figure 2-7: Runtime Error Message
Note that repairing the HLATools software does not affect your Luminex data files or database files.
1. Select the Repair option shown in Figure 2-5.
2. Click Next, then Install. The repair takes several minutes.
3. Click Finish to exit
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Uninstalling HLATools
You can uninstall HLATools either by selecting Start > Settings > Control Panel > Add or Remove Programs from
the Windows desktop, or you can use the HLATools Uninstall utility:
1. Select the Uninstall option shown in Figure 2-5.
2. Click Next, then Remove. The uninstall takes several minutes and does not affect your Luminex data files or database files.
3. If you do not plan to use HLATools further on the computer from which you have uninstalled the applications, you
may wish to remove the Luminex data files from the C:\Luminex files folder and the HLATools database files
from C:\Program Files\Microsoft SQL Server\MSSQL$ONELAMBDA.
Complete Uninstall of HLATools
In rare instances you may encounter problems running HLATools after installing a new version. These problems may
be the result of the incomplete removal of old program components and registry data during the uninstall. If this occurs,
we recommend that you perform a complete, manual uninstall of the old version and its supporting files and folders. To
do a complete uninstall:
1. Uninstall all existing instances of HLATools, the HLATools Configuration Suite and MDSE using the Windows
Add or Remove Programs utility accessible by selecting Start > Settings > Control Panel > Add and Remove
Programs.
2. Delete the HLATools folder in C:\Documents and Settings\All Users\Application Data\OneLambda. In many system installations the \Application Data folder is hidden. If you cannot find it in the \All Users folder, you can access
it by navigating to \All Users and then entering \Application Data in the Address field. Alternatively, select Start
> Settings > Control Panel > Folder Options > View > Hidden Files and Folders and choose the Show Hidden
Files and Folders option.
3. On both Windows 2000 and Windows XP, stop the Microsoft SQL Server service by selecting: Start > Settings >
Control Panel > Administrative Tools >Services > Services Local, then
•
On Windows 2000, double-click on the MSSQL$ONELAMBDA service to access the
MSSQL$ONELAMBDA properties dialog and select Stop > OK.
•
On Windows XP, select MSSQL$ONELAMBDA from the list of services and use the Stop option in the left
panel, or double-click on the MSSQL$ONELAMBDA service to access the MSSQL$ONELAMBDA
properties dialog and select Stop > OK.
When you reinstall HLATools, the MSSQL$ONELAMBDA service will also be reinstalled and started
automatically.
4. Rename C:\Program Files\Microsoft SQL Server\MSSQL$ONELAMBDA to oldMSSQL$ONELAMBDA.
5. Delete the Start\Programs\One Lambda directory from the Start menu.
6. Delete the entire folder C:\Program Files\One Lambda.
You may now install the new version of HLATools.
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Updating HLATools
When you install a new version of HLATools you might be concerned that all your existing analysis results are
preserved. To address this concern, HLATools automatically backs up your database when updating HLATools. Follow
the steps shown in Figure 2-8 to determine whether you should run Update Database, Clean Database or neither.
Figure 2-8: Updating HLATools
1. The name of the currently attached database is shown in the HLA Tools Configuration Suite Database panel
(Figure 2-9).
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Figure 2-9: SQL Server and Database Status
2. If you want to be sure that no results are lost, you can back up a database using options provided by the Backup/
Restore utility. It is recommended that you use the Backup tool since the database remains attached and available to
other users while it is being backed up. Even very large databases are typically backed up in less than one minute.
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Figure 2-10: Backup Database
3. Select the database you want to back up from the pull-down list and specify a name and path for the back-up file
using the browse feature.
4. Uninstall the old version of HLATools by selecting Start > Settings > Control Panel > Add or Remove Programs from the Windows desktop or use the removal tool provided with the HLATools Install program
(Figure 2-5).
5. Install the new version of HLATools as described above in Software Installation, p. 15.
6. At this point you have to decide whether to update the existing database, create a completely new database by running Clean Database, or maintain the current database unchanged. The Readme.txt file on the distribution CD
contains instructions on which path to follow.
7. A – Run Update Database when modifications have been made to the database or when the product nomenclature
has been updated. This preserves the results of your previous analyses.
•
Select Start > Programs > One Lambda > HLATools Configuration Suite > Clean/Update, then select
Update
•
The Update utility will take several minutes to run. Its progress is displayed in a log message window.
B – Run Clean Database when you want to create a completely new database structure. See the Clean Database
Glossary topic for details.
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Log On
In the standard installation of HLATools on a standalone workstation, any user can access HLATools without further
login. Further levels of security can be implemented to control access to the applications. The implementation of
restricted access is discussed in HLATools Security Modes, p. 149 (Appendix C).
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Chapter 3
LABType Home
LABType Home
When you launch HLATools™, the icon at the top of the list bar is the LABType Home icon shown at the right. This
icon accesses an HTML page with links to One Lambda product information in PDF format for LABType SSO and
Micro SSP product lines. The documents are included in the software distribution and reside on your local computer or
a network server, thus allowing you to access the information without going onto the Internet.
This HTML page contains links to four types of SSO reference documentation:
Product Inserts
Product insert sheets for each typing test are provided in the principal European languages required by the EU IVD
Directive. A Product Insert contains the protocol that must be followed to carry out a LABType SSO typing test.
Patient Worksheets
Patient Worksheets are used to record the reaction results for each bead in a patient sample. Each worksheet contains
the locus allele groups and their serological equivalents and positive cutoff values for each bead. The worksheets are
intended for use by laboratory technicians and HLA typing experts.
Bead Probe Information
These sheets include each bead’s probe recognition site (start and end points of various locations in HLA protein
sequence alignments that are recognized by the specific probe), allele specificities for the probe, the One Lambda
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reference test number for the bead information, and the number of true positives and negatives and any false positives
and negatives observed during the reference test. Each new revision of the Bead Probe Information sheet also includes
any changes or additions to the allele specificity list for each bead.
Resolution Limitations
Molecular genotyping is capable of much finer resolution of alleles than serological testing. This sheet contains a
listing of the alleles detected by LABType SSO testing that exhibit the same combined reaction patterns at the
serological level.
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Chapter 4
Lambda Explorer
Lambda Explorer
The HLATools™ Lambda Explorer module is the tool used to import LABScan<SuperScript>™ 100 output files into
the HLATools database.
When you first access the HLATools Lambda Explorer view, the batch files tree displays all of the Luminex files that
have been loaded into the HLATools database or which are currently available for loading.
Warning: Before you use HLATools for the first time, you must create a database structure by running the Clean
Database utility. See Running the Clean Database Utility, p. 18 for details.
Accessing Luminex .csv Files
When first launched, Lambda Explorer looks for a folder called C:\LuminexFiles. This file is the default repository for
the Luminex LABScan<SuperScript>™ 100 .csv output files which are the input files for the LTI application.
Files are displayed as follows (Figure 4-1):
•
Files in bold face type have not yet been imported into the HLATools database.
•
Files in regular type have already been imported. Attempting to re-import a file already in the database will generate a warning message.
•
Lambda Explorer parses the files to assure that their formatting is correct. Corrupt files are identified with a trash
can icon (e.g. 041204BL62.csv in Figure 4-1). Corrupt files cannot be imported in LTI.
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Figure 4-1: Lambda Explorer File Tree
To import files from a non-default folder, use the browse function by right-clicking anywhere in the file tree panel
(Figure 4-2) and select Add Folder. A conventional Find File dialog box appears.
Figure 4-2: Lambda Explorer – Adding a Folder
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Importing Luminex .csv Files
To illustrate file import, we will import one of the sample files provided with the distribution. From within the Lambda
Explorer, expand the C:\LuminexFiles folder and select ALot005.csv.
Figure 4-3: Selecting a File for Import
1
2
3
4
5
6
7
When you select a Luminex .csv file, some items of header data from the file are displayed in the main panel of the
Lambda Explorer. This allows you to review the particulars of the input file without actually importing it. The information displayed varies with the version of the Luminex software that created the data file. More extensive information
such as product lot is recorded in files made with newer versions. Figure 4-3 shows data from a Luminex 2.2 output
file.
1. Date and time – when the Luminex run was carried out (7/29/2004…).
2. Sample size – number of samples in the batch (8)
3. LX100 serial number – Luminex Reader serial number (LX10004016104)
4. Session ID – assigned at the time of the Luminex run (Aug04Workshop…). This ID is the only user-editable field
in this panel and allows the user to modify the Session ID at the time of file import. Double-clicking on the ID
enables an edit field. Edits to the Session ID affect only the version of the file imported into the database and not
the original input file, thus preserving the integrity of the original data.
5. Product designation – One Lambda product designation (RSSO1A_005) assigned at the time of the Luminex run.
Note the product designation as it will help you select the proper product/lot during the next step of the import process.
6. Lot or template name – One Lambda lot name assigned at the time of the Luminex run
(LabType A Locus Lot 005).
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7. Import – saves any edits to the Session ID and brings up the Product Select dialog box (Figure 4-4).
Figure 4-4: Product/Lot Selection
When importing Luminex output files, Lambda Explorer compares bead information in the samples with reference
values from One Lambda product data sheets and then suggests likely batch-product/lot matches as shown in
Figure 4-4. If you don’t know the lot information, select the first lot in the list. If the lot information is displayed in the
header panel, select that lot. If the match is exact, Lambda Explorer will proceed to load the Luminex file into the
database. However, if there is a mismatch, you may encounter a message like this:
Figure 4-5: Sample/Product Mismatch upon Import
A similar message appears if the samples in the batch have fewer bead types than the product.
Important Note: Please note that neither a “too many probes” nor “too few probes” message necessarily means that
the analysis cannot continue. Laboratories may customize their samples so that they do not exactly match a
One Lambda product template or a systematic laboratory error may have the same effect.
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After you OK the import or bypass the mismatch message, LTI executes five steps when importing a batch file into the
HLATools database. These are typically Trimmed Mean, Count, Normalizing, Analyzing, and Matching Patients.
•
Trimmed Mean – indicates that LTI is using as raw data the values in the Luminex data file in which the upper and
lower 5% of the readings for each bead have been discarded before the mean is calculated. The use of Trimmed
Mean data is the default setting for LTI. Other data types, such as simple Mean or Median or Trimmed Median can
be specified by the user in the HLATools Product Information Table which can be accessed by using the HLATools
Maintenance Tool (Figure 7-3). This datatype specification is made on a product/lot basis.
•
Count – imports the bead count for each bead type at each sample well. A Trimmed Count option is also available
in which the count for each bead is reduced by a specified percentage. The default value for this reduction is 10%.
The count type can be specified in the HLATools Product Information Table
•
Normalizing – normalizes the specified mean or median value for each bead; the Normalization formula can be
found in the Glossary.
•
Analyzing – determines which pairs of alleles recognized by the LABType product have combined reaction patterns consistent with the sample reaction and uses this information to construct the Reaction Pattern Histogram for
the sample.
•
Matching Patients – matches samples to patients in the database. If a Patient ID is not associated with a sample,
the sample ID will serve as a default patient ID. Thus, if a sample in a newly imported batch has the same sample
ID as a sample already in the database, and the latter sample is associated with a patient, the newly imported sample is also associated with the patient ID. This allows all of a patient’s samples to be reviewed together when you
access the patient’s analysis results in the Patient Explorer (Chapter 6). Moreover, analysis results for all samples
associated with a patient can be printed out in the Patient Report which is described on page 95. Patient matches
are not made if the samples are assigned trivial IDs, e.g. 1, 2, 3, 4.
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Chapter 5
HLATools™ LABType
Interactive
LABType
Interactive
HLATools™ LabType Interactive (LTI) compares the results of the analysis of the sample data imported in the
Luminex .csv file to the reference information contained in the One Lambda Bead Probe Information and Resolution
Limitations data sheets. Analysis results are displayed in a number of interfaces that speed and greatly simplify user
review.
Results are available at two levels of detail: Batch Analysis and Sample Analysis.
Synopsis of LTI Batch Analysis Views
The three principal views that address batch analysis tasks are shown schematically in Figure 5-1. A similar schematic
for sample-related views and subviews is shown in Figure 5-7.
Figure 5-1: Batch Analysis Views
HLATools
Explorer Bar
Batch/Sample
Tree
Main Panel
Three Tabbed Principal Views:
Batch Results Header and Table
Bead Analysis (QC Data and Sample Data)
Control Values
Batch Results
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Batch Data Analysis
You can access the LTI analysis module via the LabType Interactive icon in the Explorer Bar. The batch/sample tree
will appear similar to that shown in Figure 5-2.
The batch/sample tree contains three top-level folders. You can transfer a batch from one folder to another by selecting
the batch within the folder and using the right-click menu options.
•
Inbox – active batches (.csv session files) that are currently loaded into the HLATools database and available for
review. Sample, patient and batch data in active batches are included in reports. To prevent information from a
batch from being included in a report, move the batch into the Archive folder.
•
Deleted – inactive batches that will be permanently removed from the database the next time the Clean Database
utility is run.
•
Archive – inactive batches that you do not wish to discard. Archived batches are not removed from the database
by the Clean Database utility. You can transfer Archived batches back into the Inbox at any time.
The colors applied to the batches and samples indicate the analysis and allele assignment status:
•
Black bold face – sample contains a Matched Reaction Pair (MRP), but has not been reviewed and accepted.
•
Red bold face – sample either has a false reaction, or is empty. LTI inspects the sample reaction pattern and possible allele assignments. When the software determines that the observed reaction pattern would match that of a recognized allele if the reaction state of one (or more) bead(s) were changed from negative to positive (or positive to
negative), that bead is suggested as showing a “false reaction”. Candidate false reactions typically involve beads
with readings close to the cutoff (a bit too low or too high), so flipping the reaction state is not equivalent to ignoring or rejecting the observed data.
•
Gray regular face – sample has been reviewed and accepted.
Figure 5-2: LTI Batch/Sample File Tree
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The preliminary batch analysis results are presented in three different tabbed views – Batch Results View (Figure 5-3),
Control Values View – Positive Controls (Figure 5-5), and Bead Analysis View (Figure 5-4).
Batch Results View
The Batch Results View provides a tabulation of preliminary analysis results. You can jump directly from the table to
the Assignments View for a given sample by double-clicking on the sample’s row.
1. File header panel – the same information seen when importing the batch using the Lambda Explorer.
2. User identifier – computer’s network domain and name of currently logged-on user.
3. Filter – options to filter the entire batch according to allele occurrences in various ethnic populations. If a sample
contains an allele that is not present in the selected population, allele assignments are not made for the sample. Of
the Japanese demographic filters, Rank A filter is the most exclusionary and Rank ABC is the most inclusive.
These population filters can also be applied separately to individual samples in the Sample Assignment Subview
(right-hand panel in Figure 5-8).
4. Batch comments – annotations and comments that pertain to the entire batch.
5. Unlock All – makes all samples in the batch available for editing and changing allele assignments.
6. Auto Accept – locks all samples in the batch globally. This avoids the need to Accept the assignments for each
sample individually. Locking/unlocking individual samples is done in the Sample Assignments Subview (righthand panel in Figure 5-8). You can force LTI to reanalyze the entire batch by clicking Auto Accept, then Unlock
All. Note, however, that this will undo any acceptances that you have entered for individual samples.
7. Print – outputs a minimal summary report consisting of Sample ID, Patient ID, well location, allele assignments,
the first line of sample comments and filter status.
8. eXport – exports a summary text file containing Sample ID, Bw serotype equivalents, and the Sample Reaction
Pattern for each sample in the batch.
9. BatchResultsGrid – each row contains information about a single sample.
10. Tool tip – contains abbreviated text of the sample comments, such as:
•
False reactions – beads whose cutoffs would have to be changed are shown on the left, the resultant allele assignments on the right
•
Ambiguous calls – the type of ambiguity and the alternative calls for each allele
•
Low Bead Count – noted if the count for any bead falls below the minimum set in the HLATools Product Information table (Figure 7-3)
11. Table Layout Options Menu (abbreviated) – A full listing of the columns that can be included in the Batch
Results Grid is accessible by right-clicking anywhere in the table itself. Information that is always included by
default is indicated by an asterisk (*) in the Columns comments below. Customer-specific information is indicated
by a double dagger (‡). Customer-specific display of results is addressed in Specifying the Data Transfer File
Export Type, p. 108. Column order can be modified as discussed in Appendix A, LTI Interface Customization.
Right-click menu options
•
Save – saves the selected columns and column order
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•
Load – reloads the previous columns and column order, thus undoing any unsaved changes
•
Restore – restores the columns and column order to the HLATools default
Figure 5-3: Batch Results View
1
5
4
3
6
7
2
8
9
10
11
12
13
Row color codes
•
Blue/White – Matched Reaction Pairs (MRPs) have been assigned; i.e. the combined reaction patterns of an
allele pair match the observed sample reaction pattern. See the discussion of Allele 1/Allele 2 columns below.
All samples that have been reviewed and accepted are assigned an alternating blue or white color except for
those with ambiguous assignments.
•
Red – sample needs review, contains possible false reactions, or is empty. MRPs are possible by flipping bead
reaction states as explained in the Tool Tip discussion above (§10).
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•
Orange – ambiguous assignment which means that more than one allele assignment can be made to one or
both alleles based on the observed sample reaction pattern. The current bead combination does not provide sufficient resolution to discriminate between two or more possible allele assignments. Typical ambiguous states
are when an MRP would require a false reaction or the suggested assignments for an allele do not agree at the
subtype level.
Columns
•
Sample ID* – until a unique Patient ID is assigned to the patient, the Sample ID also serves as the Patient ID;
multiple samples in one or more batches may be associated with a single patient with a unique Patient ID.
•
BatchID* – the number assigned to the batch in the LTI database; this is not the session or batch identifier included in the Luminex .csv file.
•
Well* – well numbers and X/Y grid well location on the tray.
•
SampleID – the number assigned to the sample in the LTI database; this is not the same as the Sample ID.
•
Allele 1*/Allele 2* – allele assignments for the paternal and maternal alleles at the locus (in no particular order); if the sample indicates that the patient is heterozygous at the locus, both columns will contain a different
allele assignment; if homozygous, only the Allele 1 column will contain an entry; if no assignments can be
made, both columns remain empty. The definition of the NMDP or local code for each Allele is viewable in a
tool tip.
•
Comment* – displays the first line of the sample-specific comments and diagnostics; specifically any warning
or alert generated by the program such as “Low Bead Count” or “Low Positive Control” or notification of an
ambiguous assignment; see comment to the Tool Tip above (§10).
•
Requester – the name of the institution requesting that the sample be tested.
•
Patient ID* – part of the patient information that can be entered and edited in the Patient Information Subview; see notes to Sample ID above.
•
Lck* – checkbox displays whether the sample allele assignments have been accepted; further changes in allele
assignment, or edits to patient information or sample and patient comments can be made only if the sample is
unlocked in the Sample Assignment subview (Figure 5-8). Allele assignments that have been Locked will appear in the analysis reports as Corrected.
•
Relationship*/Family* – patient information that can be entered and edited in the Patient Information Subview.
•
A 1*/A 2* – type assignments for Allele 1 and Allele 2.
•
Sero 1*/Sero 2* – serological antigen equivalents for Allele 1 and Allele 2.
•
Bw 1/Bw 2 – Bw4/Bw6 serotype for Allele 1 and Allele 2.
•
CR* – count of Close Reactions; a close reaction is one that is just one bead reaction status change away from
yielding an MRP. See Closest Reactions Subview, p. 55.
•
MRP* – count of Matched Reaction Pairs.
•
Comment II* – user-supplied stock comment applied using the Comments pull-down menu (Figure 5-8, §12).
•
Tech1‡/Tech2‡ – usernames of preliminary and confirming reviewers.
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•
Date1‡/Date2‡ – time stamps for preliminary and confirming reviews.
•
NMDP1*/NMDP2* – definitions of NMDP or local codes used when Allele 1 and Allele 2 have been reviewed and accepted
•
A1 4Digit‡/A2 4Digit‡ – type/subtype assignments for Allele 1 and Allele 2; the assignment reflects the most
frequently observed subtype
12. Bead Analysis – accesses the Bead Analysis View which compares sample batch bead profiles with QC (reference) bead profiles. For details, see Bead Analysis View, p. 42.
13. Control Values – accesses the Control Values view which displays values for the control beads (Control Values
View – Positive Controls) and the bead counts in a sample (Control Values View – Minimum Bead Count).
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Figure 5-4: Bead Analysis View
6
1
2
3
7
10
11
4
9
12
5
8
13
16
14
Jump directly to a bead by
double-clicking on the Bead ID
18
17
15
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Bead Analysis View
The QC Data and Sample Data Bead charts allow you to observe on a bead-by-bead basis how the batch samples
compare to a representative cross-section of the general population. In both charts the bead readings for all samples are
displayed with values increasing from left to right. Clicking on the thumbnails (4, 5) displays full-sized versions of the
charts in the main panel. The False Reaction Indicator (13) in the bottom panel does not change when you toggle
between the QC Data (4) and Sample Data Bead (5) charts.
Table 5-1: Bead Analysis Color Codes
Reading
Color
Description
Very Positive
More than 110% of cutoff (assuming default 10% cutoff sensitivity)
Positive
Between 100% and 110% of cutoff
False Positive
Overrides any other positive color assignment
False Negative
Overrides any other negative color assignment
Negative
Between 90% and 100% of cutoff or bead has been used in False
Reaction assignments involving multiple beads
Very Negative
Readings much lower than 90% of the cutoff value often provide
insight into the level of background noise in the batch
1. Current Bead ID number
2. < > – use these navigation buttons to move to the adjacent bead; alternatively, jump directly to any bead in the
assay by double-clicking on the bead ID number in the bottom panel.
3. Exclude Bead from Analysis – Checking this box removes the bead from analysis. The bead will no longer be
used in making assignments such as Close Reactions, although it will still be displayed in the Reaction Pattern Histogram. In the Reaction Grid table, the columns of excluded beads are indicated by a medium gray background.
Note that removing a bead reduces the resolution of the analysis. The exclusion only takes effect if the Bead Analysis view is unlocked (17).
4. QC Data – readings were collected by analyzing 96 individuals taken from the general population and generating
profiles for each bead. By default, the Sample Data is shown in the main display when the Bead Analysis View is
accessed. Clicking on the QC Data thumbnail makes it appear in the main display.
5. Sample Data – corresponding profiles for each bead in the assay. As the number of samples in the batch increases,
the Sample Data bead profile should more closely resemble that of the general population. If the bead profiles in
the Sample Data vary greatly from the QC data, it may indicate that the samples in the batch are not representative
of the general population, or that some sort of systematic error occurred while preparing the samples or collecting
the sample data.
The Cutoff Sensitivity delta value is user-modifiable and can be changed in the HLATools Product Information
table (Figure 7-3). The default delta value is 10%. Thus, if the Cutoff Sensitivity were reduced to 5%, a reading
above 105% would count as very positive and a reading below 95% would be very negative.
6. Header – displays the name of the QC Data file or the Sample Data bead profile, whichever is selected.
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7. Y-Axis – cutoff for the bead as a percentage of the bead’s positive control value. The Y-axis is scaled vertically to
accommodate the range of the data.
8. X-Axis – sample IDs for the individuals making up the cross-section of the reference population (when QC Data is
displayed) or sample IDs (when the batch Sample Data is displayed).
9. Cutoff – default value for the bead cutoff. This value is applied globally to all samples across the batch. Cutoff values for beads can be changed locally on a sample-by-sample basis in the Reaction Pattern Histogram in the Assignments view (Figure 5-8).
10. Globally Changed Cutoff – to change the cutoff value for the bead globally:
•
clear the Locked check box (17)
•
click on the location for the new value for the cutoff; this automatically relocks the view.
To restore a globally changed cutoff to the default value:
•
clear the Locked check box
•
click on the blue default Cutoff line; the global change line will disappear.
11. Sample Specific Change Indicator – when a bead cutoff has been adjusted in the Reaction Pattern Histogram for
a specific sample, this will be indicated by an arrowhead that points in the direction of the previous setting. In
Figure 5-4, bead 59 has been changed globally as indicated by the repositioned global cutoff line, and then locally
in sample 2436212 as indicated by the upward pointing arrowhead.
12. Tool tip – mousing over a column displays the sample ID number and the normalized bead reading; double-clicking on the column opens the Assignment view for the sample.
13. False Reaction Indicator – shows the number of times that a bead has been used in an allele assignment involving
a single change in reaction status for a given bead (i.e. a Close Reaction). False Positives are bright red and False
Negatives are dark green. Changes in cutoff values are immediately reflected in this chart.
14. Y-axis – number of Positive/Negative False Reactions posited by LTI when making suggested allele assignments in
Close Reactions.
15. Y-axis – Bead ID numbers in the batch; double-clicking on a bead ID number opens the Bead Analysis view for
that bead.
16. Tool tip – dwelling over a histogram column displays the bead ID and the count of Close Reactions in which the
bead is involved; double-clicking on the column opens the Bead Analysis view for that bead. Note that Mousing
over a histogram column for one type of false reaction causes the columns for the opposite type of reaction to
become dim. This makes it possible to determine when false positives and false negatives for the same bead are
superimposed one upon the other.
17. Locked – when checked, prevents the bead from being excluded from analysis and any modifications of the cutoff
value.
18. Print – prints the histogram currently displayed in the main panel (QC Data or Sample Data).
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Control Values View – Positive Controls
The positive control values are used in the normalization calculations for the individual bead fluorescence values. The
control values are actual fluorescence readings. The formula used to normalize the bead readings can be found in the
glossary.
1. Positive Control Values – the Y-axis indicates the fluorescence value of the positive controls. There values typically range from 1000 to 4000. LTI automatically rescales the Y-axis to accommodate the range of values in the
batch. In Figure 5-5 the range of the Y-axis has been scaled down to about 2000.
2. Control Probe Symbols – the display differs according to the product being used in the analysis. Class I products
employ two positive control probes: one for exon 2 and another for exon 3. Class II products employ a single positive control probe for exon 2.
•
Green circles represent Exon 3 positive control values. These are used only in Class I products.
•
Blue squares represent Exon 2 positive control values. These are used in both Class I and Class II products.
See the Glossary for a brief discussion of Class I and Class II MHC proteins.
The minimum Positive Control Value can be set globally in the HLATools Product Information table, Figure 7-3.
The default minimum value is 1000.
3. Tool tip – mousing over a symbol displays a tooltip containing the name, value and sample ID of the positive
control bead(s) on the Positive Control Values chart.
Figure 5-5: Control Values – Positive Control Values
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2
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Control Values View – Minimum Bead Count
1. Minimum Bead Count – the Y-axis indicates the specified Minimum Bead Count for the sample or the observed
bead count if the count for any individual bead falls below the specified minimum. The Luminex 100 Analyzer
continues to log beads until the threshold has been reached for every bead or until the well is exhausted.The
Minimum Bead Count can be set globally in the HLATools Product Information table, Figure 7-3. The default minimum value is 100.
2. Bead Count Symbol – the actual bead count is indicated by a blue square.
3. Tool tip – mousing over a symbol displays the minimum bead count or the actual bead count (if less than the minimum).
Figure 5-6: Control Values – Minimum Bead Count
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3
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Synopsis of Sample Analysis Views and Subviews
There are four principal Views devoted to sample-related tasks and analysis: Assignments, Bead Analysis, Reaction
Grid and Raw Data. The Assignments view contains a main panel with five vertically tabbed subviews, a panel on the
right with two tabbed subviews for Sample Assignment (allele typing and serology matching) and Patient Information (patient demographic information), the Positive Control Values panel and the Reaction Pattern Histogram.
Tabs at the bottom access the other principal views and return to the Batch Analysis View. The views and subviews that
address sample analysis tasks are shown schematically in Figure 5-7. A similar schematic for batch-related views is
shown in Figure 5-1.
Figure 5-7: Assignments View and Subviews
HLATools
Batch/Sample
Explorer Bar Tree
Main Panel
Five Tabbed Subviews:
Right Panel
Two Tabbed Subviews:
Matched Reaction Pairs
Sample Assignment
All Alleles
Patient Information
Allele Type/Subtype
Closest Reactions
Bead Specificities
Positive
Controls
Panel
Reaction Pattern Histogram
Return to Batch Analysis and Tabs to Four Sample Views
Raw Data
Batch Analysis Assignments Bead Analysis Reaction Grid
Important Note: In the Assignments view the vertical tabs to access the five main panel subviews may be initially
hidden on small monitors or if the window is not maximized. On large monitors, an open field may appear between the
five vertically tabbed subviews in the center and the two horizontally tabbed subviews on the upper right. This extra
space has been provided to accommodate small monitors.
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Sample Results Analysis
Assignments View with Matched Reaction Pairs and Close Reactions
The Matched Reactions subview is one of a set of five vertically tabbed subviews accessible through the Assignments
view. These subviews display information about the sample and also contain tools and fields for analyzing and
modifying the sample allele assignments. Figures 5-8 and 5-9 display the Matched Reactions subview, the Sample
Assignment subview, the Control Values panel and the Reaction Pattern Histogram. Figure 5-8 shows a typical
sample with two Matched Reaction Pairs, while Figure 5-9 shows a sample with two Close Reactions. The following
notes apply to both figures.
Figure 5-8: Assignments View with two Matched Reaction Pairs
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1. Allele 1/Allele 2 – an exhaustive listing of allele assignments for the loci on the paternal/maternal chromosomes.
Selecting a cell in either column causes the Reaction Pattern Histogram (20) to highlight the part of the sample
reaction pattern that belongs to that allele (Figure 5-10).
2. False Reactions – this column contains the IDs of the bead(s) that, if their reaction state were flipped to the opposite (negative to positive or positive to negative), would allow the alleles listed in the Allele 1/Allele 2 columns to
form an MRP. The False Reactions in (Figure 5-9) each involve only a single bead, so they are considered “Close
Reactions”.
3. Code – allele assignment in the selected cell in the Allele 1/Allele 2 columns; when multiple allele share the same
reaction pattern, this is indicated by an NMPD or local ambiguity code.
4. Allele Listing – the definitions of the code (3) are displayed in this area.
5. Tabs – access four subviews:
•
All Alleles – a listing of recognized allele pairs and their serological equivalents. (See All Alleles Subview,
p. 53.)
•
Type/Subtype – a listing of all the allelic types and subtypes contained in recognized allele pairs. (See
Type/Subtype Subview, p. 54 and Type/Subtype Allele Assignments – How are … made?, p. 125.)
•
Closest Reactions – a tabulation of all Close Reactions for the current sample - i.e. reaction pattern combinations that are only one false reaction removed from yielding an MRP. (See Closest Reactions Subview, p. 55.)
The False Reactions in Figure 5-8 involve only a single bead so they also appear in Figure 5-13, Closest Reactions Subview.
•
Specificity - the allele specificities for each bead in the sample. Mousing over a bead in the Reaction Pattern
Histogram causes its specificities to appear in this subview. (See Specificity Subview, p. 56)
6. DNA – displays the allele assignments when at least one MRP has been found. When more than one MRP has been
identified, the allele codes in the left- and right-hand DNA cells combine all the Allele1 and Allele2 assignments in
the MRP table columns into a single code. In Figure 5-8 both MRPs have the same allele1 NMDP code, so this
code is shown in the left-hand cell. At the time of the analysis, there was no NMDB code that would include both
B*52AD and B*520101, so these are listed separately in two distinct MRPs. In the right-hand DNA cell, combined
B*52AD and B*520101 are represented by a local ambiguity code, B*52XX2.
7. Serological and Bw or Cw 1-Cw2 – display serological equivalents and, for B locus or C locus typing, associated
Bw4/Bw6 or Cw1/Cw2 allotypes of the HLA allele pairs shown in the All Alleles subview; these fields remain
blank if no serotype equivalent can be found or if the corresponding alleles are null alleles (e.g., BBlank or
CBlank). If the B locus or C locus allelic subtype has been identified on the molecular level but not serologically,
this is indicated by the symbol B- or C-.
8. Print – prints the OLI product name, batch name and sample-specific information contained in Matched, All Alleles, Type/Subtype and Closest Reactions subviews. The Reaction Pattern Histogram is printed on a separate page.
9. Restore – removes any user edits or assignments for the current sample from the database.
10. Locked – prevents further edits or assignment changes from being made to the sample. When this checkbox is
grayed out, the sample is not locked. To lock the sample, press Accept which enables the Locked checkbox and
checks it. A locked state for the sample is also indicated by the entries in the DNA allele assignment fields being
displayed in bold type. Locked samples are checked in the Batch Results View table (Figure 5-3).
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11. Filter – filters the current sample locally using one of the Japanese or European filters discussed above in the
Batch Results View. Allele assignments made in filtered samples are indicated by red type in the All Alleles and
Type/Subtypes subviews.
12. Comments – a short listing of frequently used user-supplied sample comments; a comment selected from the list is
appended to the sample comments contained in the field below. This entries in the list can be edited using the Comments Editor utility (see Editing the Assignment Subview Comments List, p. 107).
13. Comments Text Field – contains application-generated sample analysis messages which are also included in many
of the sample reports; this is a user-editable field – user-added comments will be included in the reports. Comments
of the following sort are displayed here:
•
Warnings – alerts of low bead count and low positive control values, when applicable
•
No False Reactions – count of the number of MRPs in the sample and the allele assignments for each MRP;
expansions of ambiguity codes are also provided.
•
Close Reactions – count and listing by bead of possible False Positive and False Negative bead reactions that
would yield additional MRPs.
•
Modified Beads – listing of beads whose cutoffs have been modified by the user.
14. Previous/Next – navigation buttons to move to adjacent samples. It is possible to jump to any sample in the batch
by selecting it in the batch/sample file tree.
15. Accept – commits the current sample assignments and comments, including any user edits, to the database and
locks the sample from further edits. Clearing the Locked check box re-enables the Accept function.
When using the option that requires that samples be accepted and reviewed by two different analysts rather than
one, the Accept button changes to read Review after the first analyst has completed his or her review. The sample
is Locked after the first acceptance has been completed and cannot be unlocked until the second review has been
completed. Changes in analysis parameters that may affect allele assignments can be made only when the sample is
unlocked.
For more on invoking customer-specific functions such as double acceptance, see Specifying the Data Transfer
File Export Type, p. 108.
16. Patient Information – patient-specific information is entered in the Patient Information subview. The same patient
information can be associated with multiple samples in one or more batches.
17. Patient – tab to access the Patient Information subview
18. Pos. Ctrl. – histogram of the raw fluorescence values for the positive control beads; Class I products type alleles in
two exons so they have two positive controls; Class II products type only one exon.
19. Arrow – collapses/expands the Positive Controls histogram; collapsing the Positive Controls panel provides more
screen space to view the Reaction Pattern Histogram.
20. Reaction Pattern Histogram – displays normalized bead readings for the sample reaction pattern. Allele assignments and alerts to conditions such as low positive control values or low bead counts are displayed in the header
field. Selecting Allele 1 or Allele 2 (or both) in the MRP table displays the corresponding assignment(s). The Reaction Pattern Histogram allows the reviewer to adjust bead cutoff values on a local basis and view the resultant
changes in suggested allele assignment. It is particularly useful for visualizing what adjustments would have to be
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made to Close Reactions or reactions with multiple false reactions to obtain an MRP. For color assignments in the
histogram, see Table 5-1, Bead Analysis Color Codes.
Reaction Pattern Histogram Symbology:
Positive reactions in Allele1
Positive reactions in Allele2
Positive reactions present in both alleles
Cutoff value
Cutoff adjusted downward, forcing a negative reaction to become positive (points toward original setting)
Cutoff adjusted upward, forcing a positive reaction to become negative (points toward original setting)
Selecting a cell in the Allele 1 or Allele 2 columns in the Matched subview causes the Reaction Pattern Histogram
to display the symbols for the beads in the sample reaction pattern that belong to that allele (Figure 5-10). It also
causes the allele assignment in the header to display the full code definition (4). Clicking Restore (9) undoes any
user-initiated changes to the bead cutoff values and restores the original assignments.
21. Y-axis – normalized bead reaction values (percent of cutoff value); see Eq. (Gl-1).
22. X-axis – bead ID numbers; control bead(s) are not included.
23. Cutoff adjustment – adjust by dragging the bead symbol vertically; the current adjustment appears in a tooltip
(Figure 5-9 only).
24. Tool tip – mousing over a bead displays a tool tip containing the information displayed in the Reaction Pattern Histogram (Figure 5-8 only).
25. Bead – the X symbol indicates the Allele 1 reaction pattern.
26. Bead – the + symbol indicates the Allele 2 reaction pattern.
27. Bead – the diamond symbol indicates that the bead reaction is common to both Allele 1 and Allele 2.
28. Bead – the downward pointing arrowhead indicates that the cutoff has been increased, with the result that the original positive has become a negative. Arrowheads point in the direction of the original value of the cutoff.
29. Tabs – return to the Batch Results Table or access the other principal sample views:
•
Bead Analysis – this view is identical to the Batch Bead Analysis.
•
Rxn Grid – a serogram-like display of alleles and their associated bead reactions and false reactions. See
Reaction Grid, p. 58.
•
Raw Data – a tabulation that includes the data shown in the tool tip (§24) plus the probe recognition site for
the bead and an abbreviated listing of bead specificities. See Raw Data Table, p. 64.
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Figure 5-9: Assignments View with Two Close Reactions
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Figure 5-10: Allele Symbols in the Reaction Pattern Histogram
Selecting a cell in the Allele 1 or Allele 2 columns causes the positive beads in that allele to display their X or +
symbols; beads in the other allele are indicated by the cutoff symbol. Selecting both cells by clicking on the pointer at
the left displays the symbols for all alleles. Beads (i.e. recognition sites) present in both alleles are indicated by a
diamond.
Matched Reaction Pairs Subview
The Matched Reaction Pairs subview lists allele pairs that match the observed sample reaction pattern. For the sake of
brevity, multiple alleles with identical reaction patterns are subsumed under an NMPD or local ambiguity code
whenever possible. This subview and its features are discussed above together with the Assignments View and the
Sample Assignment Subview. Please see Figure 5-8.
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All Alleles Subview
In contrast to the Matched Reaction Pair subview which employs NMPD or local ambiguity codes, the All Alleles table
displays all possible allele pairs down to the intron polymorphism level and lists them with their serological equivalents
(Figure 5-11). See Figure B-9 for a discussion of the method used to construct this table.
Figure 5-11: All Combinations of Both Allele Types
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7
6
1. Sero1/2 and Bw-1/2 or Cw1/Cw2 – serological equivalent and, for B locus or C locus typing, Bw or Cw allotypes
for each detected allele. To transfer the cell contents for an allele from the table to the corresponding assignment
fields, double-click on one of the Sero or Bw (or Cw) cells for that allele. Use the Restore key to revert to the original computer assignments.
2. DNA – allele assignment fields for Allele 1 and Allele 2.
3. Serological and Bw or Cw – serology and, where appropriate, Bw or Cw allotype assignment fields. Serological
equivalents displayed here will be included in the patient and sample reports when the sample has been reviewed
and accepted. The serological equivalents and Bw4/Bw6 or Cw allotype entries are editable, allowing the reviewer
to override the computer-generated assignments. To edit these values:
•
Enter new value(s) into the Serological or Bw (or Cw) edit fields.
•
Click Accept to commit the changes to the database. The emended serological equivalents will appear in the
reports, but the original values are maintained in the table.
•
The DNA assignments now appear in bold face type to indicate that the allele assignments have been reviewed
and Accepted. Allele assignments that have been Accepted are designated Corrected Typings in the reports.
When the assignments for a sample have been accepted, the sample ID in the Batch/Sample tree is displayed in
regular (non-bold) typeface.
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4. Previous/Next – navigation tools for moving to the All Alleles tables of adjacent samples.
5. Patient (demographic) data – this information can be edited in the Patient subview accessible via the Patient tab.
6. Patient – accesses the Patient Information subview.
7. Filtered types and subtypes – applying a demographic filter to the sample (outlined in red) results in a smaller
number of subtypes in Allele 1 and, consequently, different ambiguity codes in the allele assignments.
Type/Subtype Subview
This subview tabulates the allele types and subtypes for each allele. The side-by-side placement of the two lists is not
intended to imply any allele pairing. The method used to construct the Type/Subtype lists is discussed in Type/Subtype
Allele Assignments – How are … made?, p. 125.
Figure 5-12: Type/Subtype Subview
1
3
4
2
Figure 5-12 shows two type/subtype listings.
1. Unfiltered – shows all the allele assignments for the sample. The allele assignments (or in the case of multiple subtypes, the NMDP or local codes) are displayed in the column headings.
2. Filtered – applying a demographic filter to the sample may eliminate numerous subtypes. The effect of applying
the European demographic filter to the original non-filtered subtype listing is shown in red.
3. B*44ANAW – filtering the alleles results in a smaller number of subtypes in Allele 1 and, consequently, a different
NMDP code.
4. DNA – in each case the allele assignments are displayed in the DNA assignment fields.
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Closest Reactions Subview
If a Matched Reaction Pair (MRP) can be obtained by reversing the reaction state of a single bead (positive to negative,
or vice versa), that reaction pattern is known as a Close Reaction. All Close Reactions for a sample are listed in the
Closest Reactions Subview (Figure 5-13). The Closest Reactions subview shows what would happen if the cutoff
values for a bead were changed without actually doing so. The two False Reactions that appear in the Matched
Reactions subview in Figure 5-8 each involve only a single bead, so they are also listed here in the Closest Reactions
tabulation. If the False Reactions in Figure 5-9 had involved two beads or more, they would not appear here.
Figure 5-13: Closest Reactions Subview
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2
5
1. False Reactions – Close Reactions (one False Reaction) are grouped by bead and reaction type (False Negative or
False Positive). The default sorting mode of the subtables is by ascending bead number. Other sorting modes can
be specified by the user. See the Modifying Tables, p. 105, for details on table customization and modification.
2. Cells – display the suggested allele assignments that would result if the reaction status of a False Reaction bead
were reversed. Thus, if the cutoff for Bead 17 in Figure 5-13 were changed to force the reaction to be negative, the
allele assignments would be those shown in the table. If a cell in either of the Allele columns displays an NMDP or
local code, selecting the cell causes its definition to appear in the Listing to the right.
3. Pull-down list – definition of the NMDP or local code.
4. Listing – definition of the NMDP or local code, including synonymous and intron polymorphisms, and non-coding
and low expression alleles.
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5. Collapse/Expand – closes/opens all false reaction subtables simultaneously. Individual subtables can be
opened/closed by toggling the +/- sign to the left of the subtable heading. Figure 5-13 contains eight subtables.
Specificity Subview
The Specificity subview (Figure 5-14) displays the complete allele specificities for each bead in the sample.
Figure 5-14: Specificity Subview
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2
4
3
1. Bead ID – bead ID number in the batch (not the One Lambda bead identification number).
2. Specificities – complete currently known specificities for the bead.
3. Mouse over – specificities appear when mousing over the bead in the Reaction Pattern Histogram at the bottom of
the Assignments view (Figure 5-8).
4. Right click menu – text field contents can be copied to the clipboard. This is a non-editable field.
Sample Assignment Subview
This subview and its features were discussed above together with the Assignments View and the Matched Reaction
Pairs subview. Please see notes to Assignments View with Matched Reaction Pairs and Close Reactions, p. 47.
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Patient Information Subview
This subview is where the reviewer can enter patient information. Note that multiple samples may be associated with
one Patient. Information entered in the Patient Information Subview applies to all samples for a patient. Sample
information pertains only to the sample. All entry fields are optional and most require no explanation. The entry fields
are not locked until either the Match or Save operations have been carried out.
Figure 5-15: Patient Information Subview
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2
3
4
5
6
7
8
1. Patient ID – this is distinct from the Sample ID and is often different from the patient’s name for confidentiality
reasons. A Patient ID is associated with all samples belonging to the patient. Many samples can be associated with
a single patient, but only one patient can be associated with a sample. Clearing the Patient ID, then pressing Match
removes all entries from the subview.
2. Last, First, Middle – last and first names are included on results reports.
3. DOB – the DOB combo-box accesses a calendar utility that defaults to 01/01/1901; however, the field also accepts
direct entry of a date which is parsed by the calendar utility. This makes it easy to jump to any desired date.
4. Text field – comments entered in this text field are included in the Patient Reports that can be printed out from the
Lambda Reporter.
5. Match – if you enter a preexisting Patient ID in the Patient ID field, Match associates the current sample with that
patient; the preexisting patient information will be displayed here as well. Executing the Match function also
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enables the Save function. After using Match to associate a sample with a patient, be sure to Save the newly established association.
To dissociate a sample from a patient, clear the Patient ID field, then Match and Save.
6. Save – commits any edits to the database and locks the subview from further data entry, patient data in all other
samples associated with the patient are automatically updated.
7. Locked – clearing the checkbox enables the edit function.
8. Relationship/Family – these entries are also included in the Patient Summary reports.
Bead Analysis (Sample)
The Bead Analysis view is identical on both the sample and batch level. See Bead Analysis View, p. 42.
Reaction Grid
The reaction grid consists of a header panel and a tabulation of the reactions of the beads for the given sample.
Figure 5-16: Sample Reaction Pattern Grid
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The reaction grid table (Figure 5-16) shows the reaction pattern for the beads in the selected sample.
Reaction Grid Symbology and Background Colors:
•
The observed reaction pattern for the sample is shown on the Rxn row.
•
The columns of beads with observed positive reactions are indicated by a light blue background.
•
A red x on a light blue background indicates a true positive reaction.
•
A black x on a white background indicates a false negative reaction; for example, in Figure 5-16 if a positive
reaction for bead 82 had been observed, the sample reaction pattern would also include bead 82 and allele
DRB1*1109 would be part of the assignment.
•
A clear cell in the Rxn row indicates a true negative reaction; a clear cell in a column with a light blue background indicates a possible false positive reaction.
•
The top row lists the probes by number. The numbers of probes that were not used during data acquisition are
excluded for the sake of compactness in this example.
Table 5-2: Reaction Grid Background Colors
Reaction
Color
Description
Positive
Positive bead reaction if cell contains red x;
False positive if cell is empty
Observed
Reaction Pattern
Sample reaction pattern
Type color for “Golden” allele
No Reaction
No reaction
False negative if cell contains black x
Golden Allele
Allele reaction pattern is subset of sample reaction pattern
Excluded Bead
Bead globally excluded from analysis
The elements and controls in the Reaction Grid are enumerated below:
1. Sample ID – as distinguished from a Patient ID
2. (Allele1/Allele2) – suggested allele assignments; if no assignment is possible, the allele designations are left blank.
3. Allele Subtypes – display of all the alleles which define the code in the Allele column. In the case of an unambiguous allele, this will simply be the allele itself.
4. Search text boxes – used to cluster allele reaction patterns separately for Allele1 and Allele2. See Sorting by Allele
Pattern Matches, p. 63 and Reaction Grid – How is the … sorted?, p. 126.
5. < > – swaps the entries in the two search text boxes.
6. Go – redoes the clustering after an edit has been made to the entries in the search text boxes.
7. Bead IDs – you can restore the original bead order by clicking in this cell. Figure 5-21, Restoring Bead Sort Order.
8. Allele – lists all the allele specificities in the product; alleles with identical reaction patterns are represented by
NMDP or local ambiguity codes; the code definitions are shown in the right-most column of the grid (§15).
Double-clicking on any allele moves it above the Rxn row – its final position is determined by its similarity to the
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Rxn according to the criteria discussed in the FAQ topic Reaction Grid – How is the … sorted?, p. 126; doubleclicking on the Allele heading restores the original order with the sample reaction pattern (Rxn) at the top.
9. Rxn – sample reaction pattern, i.e. all the observed positive reactions in the sample; clicking in the left-most cell of
the row clusters all of the Rxn positives to the left.
10. Golden Alleles – alleles whose known reaction patterns are a subset of the observed sample reaction pattern.
Golden alleles exhibit only the positives observed in the sample reaction pattern. They are highlighted in the table
by a yellow background.
When the reaction patterns of two golden alleles can be added to yield the observed sample reaction pattern, the
junction is called a Matched Reaction Pair (MRP). In Figure 5-16, the reaction patterns of DRB1*11APWP and
DRB1*1404 together form an MRP. Similarly, the junction of DRB1*11UC and DRB1*1404 yields a second MRP.
Pairs of golden alleles yield MRPs only when their combined reaction patterns completely match the positives in
the sample reaction pattern. The combined reaction patterns of the golden alleles DRB1*11UC and DRB1*1439 do
not suffice to form an MRP, since neither shows specificity on beads 68 or 51.
11. False Positive Allele – DRB1*1404 is an example of an allele that could be part of an MRP if one of the beads in
the sample reaction pattern were a false positive, in this case, bead 10. Figure 5-17 shows another example of a
False Positive. The allele DRB1*1105 also exhibits no reaction at bead 10. If the observed reaction of bead 10 in
the sample reaction pattern were a false positive, then the junction of DRB1*1105 and DRB1*1404 would match
the revised sample reaction pattern (now minus the false positive at bead 10). Together they would form a Matched
Reaction Pair.
12. Allele Code Definition – clicking on any allele displays a full expansion of the allelic subtypes included in the
code; the subtypes are displayed in the grid header field (§3) and also in the right-most column of the table §15.
13. False Negative allele – Figure 5-18 shows an example of a False Negative allele. Allele DRB1*1109 exhibits a
positive reaction at bead 82 that is not observed in the sample reaction pattern. If the missing reaction of bead 82 in
the sample reaction pattern were due to a false negative reading (i.e., a positive reaction should have been
observed), then the junction of DRB1*1109 and DRB1*1404 would match the sample reaction pattern (which
would now also display a positive reaction) and form a Matched Reaction Pair.
LTI searches for and suggests candidate alleles for assignment by looking at so-called false reactions. These may
be either false positive and false negative alleles with known reaction patterns not too different from the observed
sample reaction pattern as seen above in Figure 5-17 and Figure 5-18. LTI first considers so-called Close Reactions
– allele combinations that are just one false reaction (positive or negative) removed from the observed sample
reaction – so that flipping the reaction state of a single bead would yield a Matched Reaction Pair. Then it looks at
suggested combinations with two false reactions, then three, and so forth.
14. Bead ID – the bead ID number provides access to several kinds of information:
•
Mousing over the bead ID number displays a tooltip with information about the bead reading. Beads 51 and 9
are examples of positive and negative reactions, respectively.
•
Clicking on a bead ID number re-sorts the Reaction Grid so that all positives for that bead move to the top of
the table and also displays the allele specificities for the bead in the header area. Figure 5-19 shows the alleles
in which bead 51 is known to display a positive reaction.
15. Allele Code Definition (not shown) – the allele code definition for an NMDP or local ambiguity code is repeated
in the right-most column of the grid.
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16. Print – prints out the reaction grid as currently displayed with definitions for the alleles on the right; due to its
width, the reaction grid should be printed out in landscape orientation, preferably on at least 11 x 17-inch paper.
Figure 5-17: Reaction Grid – False Positive
Figure 5-18: Reaction Grid – False Negative
Figure 5-19: Reaction Grid – Sorting by Bead Usage
Sorting the Reaction Grid
LTI incorporates a number of very useful sorting functions that simplify comparison and inspection of alleles and
reaction results in the Reaction Grid. When you first access the Reaction Grid, the observed sample reaction profile and
the Golden alleles appear at the top (Figure 5-16).
Clustering Positives to the Left of the Grid
You can group all the positive reactions for a given allele to the left of the table by clicking in the empty cell directly to
the left of the allele designation (Figure 5-20). Note the revised bead order in the top row of the table in ascending
numerical order: 6, 7, 11, ….
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Figure 5-20: Clustering Positive Reactions to the Left
Restoring Allele Order
You can restore the bead order to its original state by clicking in the upper left cell of the table just to the left of the
Allele column header (Figure 5-21).
Figure 5-21: Restoring Bead Sort Order
Shifting Alleles to the Top of the Grid
You can shift an allele above the sample reaction pattern in the Rxn row by double-clicking anywhere in its row. Its
final position is determined by its similarity to the observed reaction pattern. Grouping alleles together in this way
facilitates comparison with the sample reaction, making it easier to see which combinations of alleles can form a
Matched Reaction Pair. Examples of pairs of alleles shifted for this reason were seen above in Figure 5-17 and
Figure 5-18.
Restoring Allele Order
You can restore the allele listing to its original order with the golden alleles grouped at the top by double-clicking on
the Allele column header. The other alleles are displayed in descending order in terms of their closeness of match
(Figure 5-22).
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Figure 5-22: Restoring Allele Sort Order
Sorting by Bead Usage
You can also sort the Reaction Grid to see how many times a given bead was involved in allele reactions by clicking on
the bead number in first row. Figure 5-19 shows that bead 51 was involved in eight allele reaction patterns. Clicking on
the bead number or anywhere in that bead’s column also displays the allele specificities for the bead in the header
panel.
Sorting by Allele Pattern Matches
The Reaction Grid header panel contains two text entry fields that are used to enter allele designations. LTI then sorts
all the alleles in the reaction grid so that you can inspect the extent to which the specified alleles match the sample
reaction pattern. In the following discussion the alleles specified in the fields will be referred to as “First Allele” and
“Second Allele”.
Figure 5-23: Reaction Grid Sort Fields
Entering a First Allele in the left-hand sort field sorts and clusters the alleles according to the degree that their patterns
match that of the First Allele (Figure 5-24). In the first analysis, the criterion for matching the First Allele is that the
alleles share at least 75% of the positive reactions of the First Allele. The order in which the alleles are sorted is determined by this criterion and four other scores for each allele. The scoring and sorting protocol is discussed at length in
the FAQ topic Reaction Grid – How is the … sorted?, p. 126.
The alleles considered similar are sorted upwards from the specified allele in terms of decreasing similarity, with the
most similar allele being placed directly above the First Allele and the allele with only 75% similarity ending up at the
top of the grid. The sample reaction pattern appears directly below the First Allele. At this point, the remaining alleles
in the grid are ordered in terms of their similarity to the allele directly below the First Allele.
Entering a Second Allele in the other sort field moves that allele directly beneath the sample reaction pattern and forces
the remaining alleles to be reordered according to their similarity to the Second Allele. In this sorting, the alleles are
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sorted downward in order of decreasing similarity. For a more detailed discussion of the sorting methodology, see
Reaction Grid – How is the … sorted?, p. 126.
The swap button ( < > ) between the two sort fields switches the entries. See §5 to Figure 5-16.
Figure 5-24: Two-Way Sort by Allele Pattern Match
This bidirectional pattern sorting is a powerful tool for clustering allele reaction patterns. An allele with a high density
of positives at the front end often corresponds to an Allele1 (or Allele2) match, while one with a high density in the
remainder of the pattern corresponds to an Allele2 (or Allele1) match.
Figure 5-24 shows a small section of the reaction grid for a B-locus sample. Due to space constraints many rows above
and below those shown have been omitted. Similarly a large number of columns have been omitted. Many of the
omitted columns contained varying numbers of false negative reactions.
Note how this bidirectional sorting forces the alleles with high proportions of positives at the beginning of the sample
reaction pattern to cluster above the Rxn row and those with positives in the remainder to cluster right below it. This
clustering makes it easier to visualize how various combinations of alleles can yield Matched Reaction Pairs (MRPs)
and Close Reactions. In this particular sample the junction of B*1502 (Allele2) and B*0723 (Allele1) yields one of
several MRPs.
Raw Data Table
The raw data table contains bead-specific raw and normalized data. All columns in the table can be sorted alphanumerically by clicking on a column header. A secondary sort can be made by shift-clicking on a second column header.
Clicking on the Bead column heading restores the original sort order. For an example of a secondary sort, see
Figure 5-29.
The following notes refer to Figures 5-25 through 5-29.
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•
Bead – bead ID number in sample
•
Rxn – the reaction status of a bead is indicated by 8 (positive) or 1 (negative). A positive bead reading is also indicated by a light red cell color. For example, bead #18 in Figure 5-25 has a Data reading of 192% which is above the
56% cutoff for that bead. The cell color for positive control beads is always white to make them easily distinguishable from other beads with positive reactions.
•
Recognition site – these entries in Figure 5-25 contain the start and end points of the sequences that are recognized
by the probes. Also included are any amino acid polymorphisms that render the probe more specific. For details on
interpreting recognition site nomenclature, see the Recognition Site glossary topic.
Sorting the table by recognition site provides an insight into the remarkable specificity of the probes.
Figure 5-25: Raw Data Sorted by Recognition Site
Beads 18 and 28 differ only in that bead 18 specifies the substitution of a single amino acid (N, or asparagine) at
the 63rd position in the sequence. In all other respects the bead sequences are identical.
•
Raw – the mean, trimmed mean, median or trimmed median value of the fluorescence readings of the bead before
normalization
•
Data – the normalized value of the reading; this value is a percentage
•
Cutoff – percent cutoff that defines a positive reaction; when a bead cutoff has been modified from the original
value sufficiently so that the bead reaction changes from positive to negative, or negative to positive, the background color for the Rxn and Data cells for that bead changes to light purple.
Table 5-3: Raw Data Table Background Colors
Color
Description
Positive bead reaction
Negative bead reaction or
positive control bead
Low bead count; low positive control; modified
cutoff value has flipped reaction
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Figure 5-26: Minimum Bead Count in Raw Data Table
Bead counts that fall short of the threshold are indicated by a purple background in the table cell (Figure 5-27).
Figure 5-27: Low Bead Count in Raw Data Table
•
NC – negative control, which provides a measure of the background noise
•
PC – positive control; Class I products are assays for samples that contain two exons, thus there is a positive control for each exon. Since a given bead will be specific only for a site on a particular locus on one or the other exon,
different beads can have different positive controls. Class II products are assays for only a single exon. If a positive
control is low, a warning appears in the Reaction Pattern Histogram for the sample (Figure 5-8). A bead reaction
that has been changed due to a user modification in the cutoff value is also designated in the raw data table by a
light purple background color (Figure 5-28).
Figure 5-28: Low Readings in Raw Data Table
In Figure 5-28 the reading for bead #30 is near positive (right at the cutoff value of 10%) and the value for one of the
positive controls is below the default 1000 minimum.
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Figure 5-29: Raw Data Table – Double Column Sort
1. Rxn – the table in Figure 5-29 was first sorted by reaction status - with negative reactions (1) sorted before positive
reactions (8)
2. Raw – a secondary sort was then made for ascending raw bead readings
3. Bead – clicking on the ‘Bead’ heading restores the table to its original sort order.
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Chapter 6
Patient Explorer
Patient
Explorer
The HLATools™ Patient Explorer associates multiple sample results collected from among one or more batches with a
single patient. These results are assembled and collated on a patient-by-patient basis and presented for review here.
Reports of the collected sample results can be printed out in Patient Summary reports via the Lambda Reporter.
Important Note: Patient typings that have been “accepted” are designated “corrected” in some reports.
Overview
The patient information displayed here can be entered and edited in the LTI Patient Information Subview, p. 57.
1. Patient ID – see Patient Information Subview, p. 57 for details on using the Patient ID to associate multiple samples with a single patient. All assignments for patients with samples associated to their Patient IDs in this and other
active batches are listed here. Sample results for patients in other active batches are not displayed here unless the
patients are also present in the selected batch.
2. Date of Birth – non-editable field; to change the date display format, see Changing Date Formats in HLATools,
p. 107.
3. Last Name – non-editable field.
4. Comments – patient-specific comments viewable via a tool-tip.
5. +/- – expands/closes the subtable of sample information associated with each patient:
6. ConsensusAlleles 1 & 2 – the NMDP code for sample allele assignments. If the patient is homozygous at this
allele, only the first cell contains a value. Both cells are empty if no assignment can be made or if the assignments
are ambiguous.
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7. Comment – sample-specific comments; you can select the contents of the comment cell by clicking at left side of
the cell and dragging through all the text, then copy and paste it to the Clipboard in the usual manner.
8. Analysis Date – date of the allele assignment by LTI.
9. Batch Analysis Comment – a truncated version of batch-specific comments. Complete batch comments are
printed out via the Lambda Reporter.
10. Batch Name – session ID assigned at the time of the Luminex run or updated upon import into LTI.
11. Run Date – date of the Luminex run.
Figure 6-1: Patient Typings Table
1
2
3
4
8
5
6
9
10
11
7
You can modify the order of the columns of this table as discussed in Changing Batch Results Column Order, p. 106.
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Chapter 7
Reference Files and Their
Maintenance
HLATools™ LABType Interactive draws much of its reference information from three principal sources: One Lambda
product data sheets, molecular allele/serological equivalence tables, and the NMDP allele code listing. The HLATools
Configuration Suite which is installed along with the main HLATools software modules provides tools that make it
easy to review and update these three sources of reference information.
Survey of LTI Data Resource Files
To access the HLATools Configuration Suite dialog (Figure 7-1), select Start > Programs > One Lambda >
HLATools Configuration Suite > HLA Update.
From within this dialog you can perform the following functions:
•
File > Connection (menu option) – establish connections to databases on other servers; see Connecting to a Remote LTI Database, p. 138.
•
-> – the label at the top of the dialog displays the name of the computer on which the currently attached database
resides, the SQL server instance associated with the database, and the name of database in parentheses.
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Figure 7-1: Accessing HLATools Reference Data Files
Products > View
The HLATools Product Information table lists all the LABType product/lots that are included with the current release
of the software. Several columns in the table contain important editable default values that govern how batch data
associated with a given product/lot are handled by LTI and which type of data LTI uses.
Figure 7-2: Product Information Controls
The controls above the HLATools Product Information table have the following functions:
•
Tools > Global Settings (menu selection) – accesses the Global Values dialog where you can set the Minimum
Beat Count and Minimum Positive Control values for all products at once.
•
Tools > Manage (menu selection) – sorts the products in the Product Information table by descending order in
which they appear in the Lambda Explorer Product/Lot Selection pull-down menu (Figure 4-4). You can edit the
values in the Visible column to change their order of appearance in the menu.
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•
Refresh – fetches values currently stored in the database and replaces any edited values.
•
Cutoff Sensitivity – establishes the sensitivity delta percent value that distinguishes near positive (or negative)
bead readings from stronger positive (or negative) values. The default Cutoff Sensitivity is 10%. The distinctions
between the different reading ranges are illustrated in Table 5-1, Bead Analysis Color Codes.
•
Save – saves edited values to the database, overwriting previous values.
•
Cancel – undoes any unsaved edit made to the table.
Figure 7-3: HLATools Product Information
The columns in the Product Information table display the following information. Any column in the table can be sorted
alphanumerically by clicking on the column header. The sort order can be reversed by clicking a second time. Column
subsorting can be invoked by shift-clicking on additional column headers. For example, clicking first on the
ProductName column header sorts all products by catalog ID. Shift-clicking on the Lot column header then subsort all
products with identical catalog IDs by ascending lot number.
•
ProductName – the One Lambda assay catalog ID. The product name typically ends with the locus that is analyzed by the product.
You can delete a product from the table by clicking immediately to the left of the ProductName to select the entire
row and then pressing <Delete>. This action cannot be undone.
•
QABatchID – the database batch ID number for the product/lot.
•
Loci – the locus analyzed by the product.
•
Lot – the product lot number (product run)
•
Revision – revision number of the source sheet from which the product information was taken
•
DataType – the entry in this editable field instructs the software which raw data from the Luminex output file to
use for normalizing the bead readings. Trimmed Mean is the default. For details and examples, see the Mean and
Median Glossary topic.
•
CountType – the entry in this editable field instructs the software which count data from the output file to use. The
full count is the default. See the Count Type Glossary topic.
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MinBeadCount – a different minimum value can be set for each product/lot. For example, if a product/lot contains
a smaller number of one or more beads than normal (the default is 100), this value can be adjusted accordingly to
prevent the samples from being rejected during analysis. The value for MinBeadCount can also be changed globally using the Tools > Global Settings menu option.
•
MinPCData – the minimum Positive Control value. The default is 1000. Only one value for MinPCData is specified even with products that have two positive controls, such as lots for B locus analysis. The value for MinPCData
can also be changed globally using the Tools > Global Settings menu option.
•
Visible – indicates the priority ranging from 0 to 10 in which a product appears in the Product Select drop-down
menu when importing a sample batch into the LTI database from within the Lambda Explorer.
•
0 removes the product from the suggestions in the drop-down menu; see Figure 4-3, Selecting a File for Import. To reduce the clutter in the drop-down menu that would be caused by including out-of-date products, set
this value to ‘0’. However, since you may need to review LABType analyses that were based on earlier product/lots, it is recommended that you not delete out-of-date products from the Product Information Table.
•
10 is the highest priority and causes the product catalog lot to appear at the top of the dropdown list. Other
numbers from 1 to 9 can be assigned to force the order in which products appear. Products with identical priority numbers are displayed in the dropdown list in the order they appear in this table.
Figure 7-4: Bead Data in the Product Information Subtable
Expanding the product row by clicking on the + at the left side of the table displays a subtable (Figure 7-4) containing
specificity data for each bead. Note that all fields in these subtables are editable.
•
BeadID – ID number of sample bead
•
PosCtrlBead – ID number of the positive control bead whose control value is used to normalize the reading for the
sample bead
•
AbbreviatedSpecificity – a compact version of the complete list of alleles that is presented in the CompleteSpecificity column. The abbreviated specificity list is used in many of the output reports. The specificities are abbreviated using these rules:
•
Allele families – the entries for a family are terminated by a semicolon
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•
Alternation sign – the forward slash symbol ( / ) designates an alternative: B*1542/44 means subtypes
B*1542 or B*1544.
•
Range sign – the tilde ( ~ ) designates a range of allelic subtypes: B*1542~44 means subtypes B*1542,
B*1543 and B*1544. When allelic subtypes are included in a range, only the first four digits in the designation
are significant: all digits signifying synonymous, intron and null or low expression polymorphisms are disregarded. Thus B*1542~44 could also be the abbreviation for the series B*1542, B*154201, B*15420101,
B*154202, B*1543N and B*1544.
Because information is lost by abbreviating the specificities, LTI uses the complete specificity listing when
matching allele pairs with the observed sample reaction pattern. For a discussion of allele designation, see Alleles –
How are they designated? in Appendix B.
•
RecognitionSite – probe recognition site(s); these entries describe the site(s) on the locus that are recognized by
the probe on a given bead. For details on how to interpret these entries, see the Recognition Site glossary topic.
•
ProbeID – OLI probe identifier.
•
CutOff – the minimum value of the normalized bead reaction that is considered positive; editing this value for a
particular bead will change the cutoff in all analyses of batches using this product.
•
CompleteSpecificity – a complete listing of all allelic subtypes to which the probe sequence on the bead will bind.
Products > Update
The HLATools installation includes a utility that lets you update the LABType product information used by LTI. This
information is the same as that contained in the Bead Probe Information and Resolution Limitations data sheets for a
given LABType product. A sample of this information is shown in Figure 7-4.
To update information for a LABType product:
1. Access the listing of new One Lambda products at:
http://download.onelambda.com/pub/tray_info/Windows/HLATools/Labtype_Interactive/
2. The product filename must have a .LabTypeProduct extension as shown below. Products designated by an R
revision number have been retired. If you do download a retired product, it will appear in the HLATools Product
Information Table, but it will not appear in the Lambda Explorer Product Select pull-down listing (Figure 4-4)
since its Visible attribute is automatically set to 0. (See discussion on page 74.)
Figure 7-5: Selecting a One Lambda LabType Product to Update
3. Click on the filename hyperlink to launch the updating utility. A conventional Windows File Download dialog will
appear. To download the new product information directly into the LTI database, select the Open option. An Open
With dialog will prompt you to select an application to carry out the download operation. Select GenProd.
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Figure 7-6: Downloading One Lambda Product Information
4. One of two log messages will appear:
•
If the selected revision of the product information is already in the LTI database, a message will confirm the
fact, and the window may be visible for only a second or two. The existing version will not be replaced.
•
If the product is not present or needs to be updated, the download should take about fifteen or twenty seconds.
5. Alternatively, you can Save the file to your computer. You might choose the Save option if you want to update the
product information at a later date or if you need to transfer the data file to a second computer that is not attached to
the Internet.
When you Save a file, please note the extension associated with the saved file. If the updating utility is correctly
installed on the computer and the file extension is correct, the File Download dialog should display the icon shown
in the circle in Figure 7-6. You can load the updated product information into the database just by double-clicking
on the saved file.
To confirm that the new revision of the product information has been imported into the LTI database, launch the
HLATools Configuration Suite and select Products > View to access the HLATools Product Information table. The
newly updated product will appear in the bottom row. For details on this table, see HLATools Product Information,
p. 73.
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Important Note: When you Save a file, please note the extension associated with the saved file. The internet browser
security settings on some computers may suppress the .LabTypeProduct file extension when you download the file
although the extension does appear when you navigate to the file on the One Lambda ~LabTypeInteractive/ webpage
(Figure 7-5). When these security settings are in effect, the.LabTypeProduct file extension is replaced with a generic
.doc extension with the result that your operating system cannot associate the file with the updating utility and it will
not be able to load the file information into the LTI database. Save the file to a location on your local computer and add
the file extension to the filename. When you select the Product > Update option and browse to the folder containing
the new LABType file(s), the updating utility will be able to recognize the files. The utility will update your LTI
database with the files you select by checking the checkbox to the left of the filename (Figure 7-7).
Figure 7-7: RSSO Files with LabTypeProduct Extension
NMDP > View
This option displays a listing of NMDP allele codes and their subtypes taken from the NMDP Research web site. After
the first update, the revision date of the code listing is displayed above the table. Before the first update, the legend will
state “Unable to retrieve”.
This table lists NMDP codes and their numeric allelic subtype definitions. The LTI table is initially presented using a
numerical subtype sort (see inset in Figure 7-8). By clicking on the “Code” column heading, you can re-sort the table
so that the allele codes are displayed alphabetically. The NMDP allele codes are curated by the NMDP Research
Program. The current version may be found at http://www.nmdpresearch.org/HLA/numeric.html.
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Figure 7-8: Navigating to an NMDP Code in the NMDP Allele Code List
The NMDP Information View controls are identical in function to those in the Product Information View:
NMDP > Update
This tool fetches the latest revision of the Code List directly from the NMDP Research website or from a locally stored
file.
Figure 7-9: Updating the NMDP Allele Code List
Serological > View
Molecular genotyping is capable of detecting allelic differences down to the single nucleotide or single amino acid
level. However, for some types of transplantation procedures, especially those using cadaveric organs, allelic
genotyping is not possible. In such cases, expediency requires serological tissue matching. Many users may find it
useful to relate the results of the LTI allele analysis to their serological equivalent.
This option accesses the Serological Information table where you can view, edit and export the contents of the Allele to
Serological Equivalent table (Figure 7-10). The revision of this table current at the date of release is provided on the
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distribution CD. After the first update, the revision date of the code listing is displayed above the table. If the
Serological table has not been updated since the time of installation, the Serological Revision Date label will state that
the application is “Unable to retrieve” the revision date.
All of the cells in the table are editable fields to allow user updating and annotation.
Figure 7-10: Navigating to an Allele in the Serological Equivalents Table
The columns in the serological equivalents table contain the following information:
•
Name – the allele type and subtype assigned by OLI HLA analysis.
•
SerologicalEquivalent – many alleles have identified serological equivalents. These equivalents are displayed in
this table. Null alleles, i.e. alleles whose molecular variations are not expressed as changes in surface antigens, are
indicated in the Serological Equivalent Table as *Blank, where * represents a serological type. Allelic types that
have been identified on the molecular level but not serologically are indicated as *-.
•
BestGuess – this column is currently unused.
•
Bw4/Bw6 or Cw1/Cw2 – the Bw4/Bw6 and Cw1/Cw2 systems are biallelic antigen systems closely associated
with HLA-B and HLA-C.
The Allele to Serological Equivalent controls are identical in function to those in the Product Information view with
one addition:
•
Export XLS - exports table in Excel-compatible Comma Separated Value format. Because the file includes Excelcompatible metadata, it cannot reimported into the LTI database.
•
Export CSV - exports table in Comma Separated Value format that can be reimported into the LTI database.
Serological > Update
Updates the Serological Information table by accessing a serological equivalents file stored locally or on an internet
website.
Filter > View
The Filter View and Update features are currently under development.
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Chapter 8
HLATools™ Lambda
Reporter
Lambda Reporter
The HLATools™ Lambda Reporter provides tools for generating customized reports. Report formatting options
include:
•
Customization of header and footer information
•
Inclusion of your laboratory’s logo in the header in two different size formats
•
Five categories of predefined reports or report templates. You can edit many of these predefined templates and save
the customized templates that you have created. The report template categories are:
•
Patient Reports
•
Batch Reports
•
Sample Reports
•
•
•
Combined Sample Report
•
Sample Report
Summary Reports
•
Batch Results Report
•
Catalog Information
•
Data File Summary Report
•
Database Information Report
•
NMDP Allele Code Report
•
Patient Typing Summary Report
•
Serological Equivalent Report
Special Reports
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ABDR by Batch Report
•
ABDR by Batch Report (Expanded)
•
ABDR by Batch Report (Range)
•
Allele Group Frequency Report
•
Allele Query Report – <under development>
•
Reaction Assignment Report – NIH score for each sample
•
Swiss Lab Report – (.csv file intended for export only)
The Lambda Reporter allows the user to create reports with customized header and footer information, generate reports
in which the patient results are ordered according to a variety of sorting criteria, and group analysis results for one or
multiple patients from different test batches.
For overviews of the contents of the stock template formats, see the corresponding report description in Chapter 9,
HLATools™ Legacy Reporter.
Laboratory Report Header Information
Laboratory-specific information used by the Lambda Reporter is entered directly from the Lambda Reporter or using
the HLATools Configuration Suite.
Figure 8-1: Laboratory Header Information
To replace the One Lambda company icon shown in Figure 8-1 with your own laboratory, company or institutional
icon:
•
Select HLATools > Lambda Reporter
•
Select the Logo tab in the Search panel (Figure 8-2)
•
Select the desired aspect ratio for the new logo. Best results are obtained if the logos are the same sizes as indicated on the Logo panel: 170px x 27px for the wide logo and 38px x 29px for the small logo. If you use a long
logo, the text shown in Figure 8-1 shifts accordingly to the right.
•
Browse to the folder where the logo graphic resides. Recommended file formats are .bmp or.jpg.
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Figure 8-2: Importing a Custom Logo
Entering Laboratory Information
To enter your laboratory information, select Start > Programs > One Lambda > HLA Tool Configuration Suite >
Configuration > Lab Info.
Figure 8-3: Lab Info Interface
The Lab Info interface has several controls or fields that warrant discussion.
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•
Refresh – restores all values to those currently saved in the database.
•
LabID – can be used to identify multiple laboratories or departments within the same institution.
•
Save – saves all newly entered values or changes into the database.
•
Cancel – discards all newly entered values or changes without saving them into the database.
The other fields are self-explanatory and do not require elaboration.
Using Lambda Reporter Filters
Selecting Samples
The Lambda Reporter provides filters that allow you to specify very precisely which samples will be included in or
excluded from your reports. To illustrate these filters, we will create a Patient Report that contains several samples.
1. Select Patient Reports in the bottom panel of the Lambda Reporter sidebar to access a panel containing patient
report search options (Figure 8-4).
Figure 8-4: Patient Reports Search Options
2. Note that you can select Patient results from archived batches (Archive) as well as active batches (Inbox). In this
example we will select both options.
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3. By default, the Reporter matches up all patients from all batches (Figure 8-4). We can use the Patient ID pull-down
list to narrow the search to a single patient. The pull-down list contains the names of all the patients in the LTI database. In this case we will use a wildcard (*) to find all the patients (Figure 8-5) whose Patient IDs start with the
numerals “243613” and later select several for inclusion in our report. After entering the Patient ID search string,
we press the Search button at the bottom of the panel.
Figure 8-5: Patient Search Options
4. The search results are displayed in the Filters panel (see side tab in Figure 8-5). You can include one or more
Patients in a report by checking the Include boxes on the left. A number of the most important patient attributes
such as Gender, Race, etc. can be displayed in the Filters panel.
This table is fully sortable. You can sort on any column by clicking on the column heading to the left of the filter
cone icon, and you can perform a subsort by shift-clicking on a second column.
5. The Patients selected in the Filters table automatically appear in the Sample(s) to Report On panel immediately
below. The default sorting order for this table is by well position, although you can sort the samples by the contents
of any column in the table. In Figure 8-6 the two samples have a low bead count, so no allele assignment has been
made.
Figure 8-6: Samples to Report On
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6. Further filters for searching on the values in each column are contained in a right-click menu that can be accessed
by clicking directly on the filter cone icon (Figure 8-7). The right-click menu lists each item in the column, plus
options for selecting all items (All), items with a value (NonBlanks) or no value (Blanks).
Figure 8-7: Further Sample Table Column Search Filters
7. The right-click menu of Figure 8-7 also contains a Custom option that accesses a dialog with advanced filter criteria (Figure 8-8). With the Custom filter option you can create very specific searches for including or excluding
samples. In Figure 8-8 two conditions are applied to the search: the Patient ID must be “greater than” Sample–B4
and “less than or equal to” Sample–B7.
Note that a code representation of the selected search filter appears in the text field at the bottom of the dialog box.
(Figure 8-8).
Different sets of appropriate search operands are available for filtering on each row.
Figure 8-8: Advanced Search Filters
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8. You can preview the reports for the selected samples in the specified format by clicking on Preview in the Report
selector pane (see bottom of Figure 8-4). Figure 8-9 shows part of a Patient report.
Figure 8-9: Patient Report for One Sample
9. The Filter panel also contains a Report Summary subpanel (Figure 8-10) that can be toggled via the Hide/Show
Report Summary checkbox in the Sections tab of the Search subpanel (Figure 8-11). You must first click the
Edit button to enable the checkboxes in the Sections tab.
Figure 8-10: Report Summary Subpanel
Checking Show ‘Report Comment’ on Report includes the report-specific user comments in the report
(Figure 8-15).
The lower text field contains general instructions about the current type of report being generated. This text does
not appear in the report.
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Creating a Custom Report Template
The easiest way to illustrate how to create a custom report template is by modifying a stock template.
Figure 8-11: Sample Report Default Sections
In the Lambda Reporter:
•
Select the report category Sample Reports at the bottom of the Report Selection panel.
•
In the Search panel, select the Sections tab, then Edit. The sections included by default in the existing Sample
Report are checked and in bold-face type (Figure 8-11). An actual report containing these default sections is
shown in Figure 8-13.
•
When you have included or excluded the desired sections in the template, Save the template to the folder containing the templates of that report type. You will be prompted to rename your custom template so you do not
write over the stock template. Your new report template is added to the listing of reports of that type
(Figure 8-12).
•
Creating templates for other types of reports differs only in minor details.
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Figure 8-12: Batch Reports Listing
Figure 8-13: Default Sample Report Default
1
3
2
4
Report Sections
The sections available for inclusion in a report are described below. The default sections of a Sample Report are shown
in Figure 8-13. Other types of reports contain differing collections of default elements.
•
Lab Information (1)
•
Patient Information (2)
•
Batch Information (3)
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Short Comments (4)
Long Comments (Figure 8-14) include:
•
MRP count (5)
•
NMDP code definitions for MRP alleles (6)
•
CR count (7)
•
CR False Positive and False Negative bead numbers (8)
•
Review History (9)
•
Hide/Show Report Summary – when shown, report summary includes user-supplied report-specific comments (Figure 8-15-10) that are entered in the Report Summary field of the Filter panel (Figure 8-10)
•
Reviewer Information – names of sample reviewer and approver with timestamps when two-reviewer option
is in use (Figure 8-15-11)
•
Warning Messages (Figure 8-15-12)
•
Rxn Pattern – bead NIH scores (Figure 8-15-13)
•
PC/Beads Graph – Reaction Pattern Histogram (Figure 8-16-14) with Positive Control Bead reading
(Figure 8-16-15).
•
New Page After – creates page break; each report prints on a new page
Figure 8-14: Sample Report Long Comments
5
6
7
8
9
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Figure 8-15: Non-Default Sections of a Sample Report
10
11
12
13
Figure 8-16: Sample Report Positive Control and Reaction Pattern Histogram
14
15
Specifying Report Paper Size
The default report paper size is normally determined by the Windows OS language specification. The report paper size
has U.S. Letter dimensions when U.S. English is specified in the Regional and Language Options Windows control
panel or A4 size when any other language is specified. However, the paper size local default can be overridden in the
Lambda Report Paper Size tab (Figure 8-2).
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Chapter 9
HLATools™ Legacy
Reporter
Legacy Reporter
The HLATools™ Lambda Reporter provides tools for generating five categories of preformatted reports in either U.S.
standard (8½ x 11") or A4 (210 x 297 mm) sizes. Templates for the preformatted reports in both sizes reside in the
C:\Program Files\One Lambda\HLATools\Reports or ~\ReportsA4 folders. The logo which appears in preformatted reports is located in the file C:\company_logo.bmp and should conform to the logo dimensions cited in
Laboratory Report Header Information, p. 82.
HLATools automatically selects the appropriate paper size according to the Regional and Language Options settings in
the Windows Control Panel. If you wish to use the non-default paper size, you can override the automatic setting as
described in Specifying Report Paper Size, p. 91.
Preformatted Reports
The Lambda Reporter generates five categories of preformatted reports which are available in US Standard and A4
paper sizes as discussed above.
To use a preformatted report:
1. Launch the Lambda Reporter. By default the Lambda Reporter displays the Filters view.
2. Select Report Category > Report Type. (e.g. Batch Reports > Batch Report Sorted by Sample).
3. Invoke the appropriate filter(s) from the pull-down lists as shown Figure 9-2:
•
Batch ID
•
Sample ID
•
Patient ID
•
Allele
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•
Catalog/Lot
4. Press Report. The report is automatically displayed in the Reports view.
5. If the report is:
•
For print, use the printer icon in the tool bar; many print reports can also be exported;
•
For export only, use the envelope icon to the right of the printer icon to access a standard Save File dialog
(Figure 9-1).
Export formats include .txt., .pdf, .xls, and .doc file formats.
Figure 9-1: Report Print and Export Functions
Patient Reports
1B – Patient Summary Report Filtered by Batch
The report contains:
•
Allele assignments for all samples associated with each patient in the selected batch; allele assignments for samples
not associated with a patient are also included. NMDP codes are expanded to four digits plus N or L suffix.
•
Analysis date for each sample
•
Corrected (accepted) allele assignments are printed in a separate column
2A – Patient Report
A combined sample analysis report of all samples associated with the specified patient. Multiple samples in a single
batch or samples from different batches may be associated with a single patient. The report includes:
•
Full patient data
•
Batch/product lot information and locus type
•
Sample draw, test and analysis dates
•
Batch comments
•
Corrected (accepted) typing, if any
•
Reaction assignment (NIH 8/1 single-row format)
•
Computer assignment combining all MRP designations into a single NMDP code pair, if applicable
•
Generic allele descriptions with complete listing of types and subtypes for each MRP
•
Listing of close reactions
•
Expansion of NMDP codes for computer allele assignments
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Figure 9-2: Report Filter Interface
Batch Reports
3A – Batch Report
This report format includes as many samples as possible on each page.
•
No patient data
•
Batch/product lot information and locus type
•
Test and analysis dates for each sample
•
csv filepath
•
Batch comments
•
Corrected (accepted) typing, if any, for each sample
•
Reaction assignment (NIH 8/1 single-row format)
•
Computer assignment combining all MRP designations into a single NMDP code pair, if applicable
•
Generic allele descriptions with complete listing of types and subtypes for each MRP
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Listing of close reactions
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Expansion of NMDP codes for computer allele assignments
3B – Batch Raw Data Abbreviated Specificity Report
•
No patient data
•
Batch/product lot information and locus type
•
Draw, test and analysis dates for each sample
•
csv filepath
•
Batch comments
•
Corrected (accepted) typing, if any, for each sample
•
Reaction assignment (NIH 8/1 single-row format) is included in raw data table
•
Computer assignment combining all MRP designations into a single NMDP code pair, if applicable
•
Generic allele descriptions with complete listing of types and subtypes for each MRP
•
Listing of close reactions
•
Expansion of NMDP codes for computer allele assignments
•
Tabulated data for each bead in the sample, including:
•
Bead number
•
Reaction status (1 or 8)
•
Recognition site with amino acid mutation
•
Raw data and normalized value for bead reaction
•
Cutoff percent
•
Values for negative and positive control(s)
•
Bead count
•
Abbreviated allele specificities for each bead
3D – Batch Report Sorted by Patient
This report omits any samples not associated with a patient ID. All associated samples are listed under a patientspecific data header. Draw dates for individual samples are not included.
•
Patient data (name, gender, etc.) and patient-specific comments
•
Batch/product lot information and locus type
•
Draw, test and analysis dates for each sample
•
csv filepath
•
Batch comments
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Corrected (accepted) typing, if any, for each sample
•
Reaction assignment (NIH 8/1 format) is included in raw data table
•
Computer assignment combining all MRP designations into a single NMDP code pair, if applicable
•
Generic allele descriptions with complete listing of types and subtypes for each MRP
•
Listing of close reactions
•
Expansion of NMDP codes for computer allele assignments
3E – Batch Report Sorted by Sample
This report contains only sample-specific data. Draw dates for individual samples are not included.
•
Batch/product lot information and locus type
•
Test and analysis dates for each sample
•
csv filepath
•
Batch comments
•
Corrected (accepted) typing, if any, for each sample
•
Reaction assignment (NIH 8/1 format)
•
Computer assignment combining all MRP designations into a single NMDP code pair, if applicable
•
Generic allele descriptions with complete listing of types and subtypes for each MRP
•
Listing of close reactions
•
Expansion of NMDP codes for computer allele assignments
3F – Combined Batch Report Sorted by Sample
This report contains data for each sample in order assembled from each batch in which the samples appear. Draw dates
for the samples are not included. The csv filepath in the report header is the one selected in the Batch ID filter
(Figure 9-2). The filepath for each batch is also reported.
•
Batch/product lot information and locus type
•
Test date for each sample
•
csv filepath
•
Batch comments are omitted
•
Corrected (accepted) typing, if any, for each sample
•
Reaction assignment (NIH 8/1 format)
•
Computer assignment combining all MRP designations into a single NMDP code pair, if applicable
•
Generic allele descriptions with complete listing of types and subtypes for each MRP
•
Listing of close reactions
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Expansion of NMDP codes for computer allele assignments
3G – Combined Batch Report Sorted by Patient
This report contains data for each patient assembled from each batch in which an associated patient sample appears.
Draw dates for the samples are not included. The patient information for each patient includes Patient ID, name,
gender, birthdate, race and blood typings.
•
Batch/product lot information and locus type
•
Test date for each sample
•
csv filepath
•
Batch comments are omitted
•
Corrected (accepted) typing, if any, for each sample
•
Reaction assignment (NIH 8/1 format)
•
Computer assignment combining all MRP designations into a single NMDP code pair, if applicable
•
Generic allele descriptions with complete listing of types and subtypes for each MRP
•
Listing of close reactions
•
Expansion of NMDP codes for computer allele assignments
Sample Reports
3C – Sample Report Filtered by Batch
This brief report contains data for each sample in the selected batch. The test date, name of the tester, csv filepath and
batch comments are included in the header. The report for each sample includes
•
Batch/product lot information and locus type
•
Corrected (accepted) typing, if any, for each sample
•
Reaction assignment (NIH 8/1 format)
•
Computer assignment combining all MRP designations into a single NMDP code pair, if applicable
•
Generic allele descriptions with complete listing of types and subtypes for each MRP
•
Listing of close reactions
•
Expansion of NMDP codes for computer allele assignments
4A – Sample Raw Data Abbreviated Specificity Filtered by Batch
This report contains raw data for the specified sample in a single batch.
•
Batch/product lot information and locus type
•
Draw, test and analysis dates for the sample
•
Session ID and csv filepath for the batch
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Batch comments
•
Corrected (accepted) typing, if any, for the sample
•
Reaction assignment (NIH 8/1 format) is included in data tabulated table
•
Computer assignment combining all MRP designations into a single NMDP code pair, if applicable
•
Generic allele descriptions with complete listing of types and subtypes for each MRP
•
Listing of close reactions
•
Expansion of NMDP codes for computer allele assignments
•
Tabulated data for each bead in the sample. Note in the table footer indicate whether a bead’s positive control or
count are below the minima specified in the HLATools Product Information table
•
Bead number
•
Reaction status (1 or 8)
•
Recognition site with amino acid mutation
•
Raw data and normalized value for bead reaction
•
Cutoff percent
•
Values for negative and positive control(s)
•
Bead count
•
Abbreviated allele specificities for each bead
4B – Combined Sample Analysis Data Report
This report contains the test results for all samples with the specified sample ID in all active batches (i.e. batches in the
LTI Inbox folder).
•
Patient information for associated sample. (Only one patient can be associated with a given sample.)
•
Batch/product lot information and locus type for each batch
•
Draw, test and analysis dates for each sample
•
Session ID and csv filepath for each batch
•
Batch comments for each batch
•
Corrected (accepted) typing, if any, for each sample
•
Reaction assignment (NIH 8/1 format) is included in data tabulated table
•
Computer assignment combining all MRP designations into a single NMDP code pair, if applicable
•
Generic allele descriptions with complete listing of types and subtypes for each MRP
•
Listing of close reactions
•
Expansion of NMDP codes for computer allele assignments
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4C – Sample Raw Data Abbreviated Specificity Report
This report contains the same information as report 4A. However, whereas 4A contains information only for the
specified sample in a single specified batch, this report contains the information for all samples with the specified
sample ID in all active batches.
4D – Sample Abbreviated Specificity Report
This report contains information identical to 4C, with the exception that the tabulated raw data contain only the
following three entries:
•
Bead number
•
Reaction status (1 or 8)
•
Abbreviated allele specificities for each bead
4E – Sample Complete Specificity Report
This report contains information identical to 4C, with the exception that the tabulated raw data contain the complete
(i.e. non-abbreviated) allele specificities.
99A – Sample Report with Chart Filtered by Batch
This report contains the following information for each sample in specified batch plus a Reaction Pattern Histogram:
•
No patient information
•
Batch/product lot information and locus type for the batch
•
Draw, test and analysis dates for each sample
•
Session ID and csv filepath for each batch
•
Batch comments
•
Corrected (accepted) typing, if any, for each sample
•
No reaction assignment (NIH 8/1 format)
•
Computer assignment combining all MRP designations into a single NMDP code pair, if applicable
•
Generic allele descriptions with complete listing of types and subtypes for each MRP
•
Listing of close reactions
•
Expansion of NMDP codes for computer allele assignments
•
Normalized Bar Graph (Reaction Pattern Histogram) for each sample
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Summary Reports
1A – ABDR by Batch Sorted by Well Position
This report returns analysis exports for A, B, Cw, DQ and DR loci in Excel spreadsheet format. Because of the width of
the spreadsheet the preview will only display a small part of the report. You must open the exported file in Excel to see
all the information. The report includes:
•
Sample ID
•
Patient Name
•
Draw, test and analysis dates for each sample
•
Catalog/Lot number
•
A, B, Cw, DQ and DR assignments using NMDP code and numeric expansions of NMDP code
•
Accepted/Not Accepted status
1C – Batch Results
This report contains all of the allele assignments and numeric expansions for the samples in the specified batch. Information includes:
•
Session ID
•
Product/Lot for batch
•
Sample IDs
•
Well locations
•
Allele assignments and numeric expansions for each sample
•
Associated patient and kinship information
•
Accepted/Not Accepted status
•
Number of Close Reactions
•
Number of MRPs
5B – Allele Query Report
This report returns all of the samples in which the specified allele has been detected. Only “accepted” samples are
included in the report. The report includes for each sample:
•
Catalog/Lot number
•
Draw, test and analysis dates
•
Patient data for the sample
•
Reaction assignment
•
Corrected typing
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6A – Serological Equivalent Table Report
This report contains the same information as the Allele to Serological Equivalent table that can be accessed using the
HLATools Maintenance tool. See Survey of LTI Data Resource Files, p. 71 and Figure 7-10.
6B – NMDP Allele Code Data
This report contains the NMDP code numeric expansions sorted alphabetically. The codes can be accessed using the
HLATools Maintenance tool. See Survey of LTI Data Resource Files, p. 71 and Figure 7-8.
6C – Batch Data File Summary Report
This report contains a history of batch operations since HLATools was installed on the local computer. The following
information is provided for each batch:
•
Database batch ID number
•
Batch session ID
•
Serial number of Luminex machine used to analyze the batch
•
Luminex run date and operator
•
One Lambda product/lot used
•
Batch status (active, deleted or archived)
•
Batch filepath
6E – Catalog Information
This report contains product information for the selected LABType product.
•
Product Catalog Number
•
Lot Number
•
Applicable Product Insert Revision Number
•
Data Type employed (Mean, Trimmed Mean, etc.)
•
Count Type employed (Count or Trimmed Count)
•
Minimum Bead Count setting
•
Minimum Positive Control value
•
For each bead
•
Bead number
•
Associated positive control bead
•
Abbreviated specificity
•
Recognition site with amino acid mutation
•
Cutoff value (%)
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Complete specificity
Special Reports
5A – Allele Group Frequencies
This report contains tables and accompanying graphs of the alleles detected and their phenotypic and genotypic
frequencies in the HLA A, B, Cw, DQ, DP, DR loci analyzed using One Lambda products and LabType Interactive.
6D – Database Table Information
This table contains information about database contents and usage.
The three columns on the first page of the report list:
•
Run dates – (grouped by week) of the batches currently in the database. These numbers include both sample and
QA batches. The QA batches are used by LTI to construct the QA bead profiles. (This column currently has no
header.)
•
Batch – number of batches run during that week
•
Sample – total number of samples in the batches
•
Inbox/Deleted/Archived – current disposition of the user’s sample batches in the LTI database
•
QA Batch – number of QA batches used to construct the QA bead profiles
The information tabulated on the second page is of diagnostic interest to the database administrator.
99B – ABDR by Batch Sorted by Well Position
This report is identical to 1A, except that it returns results only for Allele1. It also includes serotypic equivalents for the
assigned allele.
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Appendix A
LTI Interface Customization
HLATools™ LABType Interactive interfaces contain a number of user-customizable features that are not immediately
apparent to the beginning user. Some of the more useful features are described in this Appendix.
Modifying Tables
The appearance and functionality of many tables in LTI are user-editable. By rearranging the column headers, you can
make the table present the data in different ways that may be more suitable for the way you work.
Changing Sort Order or Grouping
The Closest Reactions subview is an example of a table that can be reconfigured to change the way the entries are
sorted and grouped. By default, the entries are sorted by bead number (Figure 5-13). To reconfigure the table so that it
sorts the False Reactions by Allele1 candidates, drag the False Reactions header from its position at the top of the table
and drop it beside the Allele2 header. Then drag the Allele1 header and drop it at the top. The reconfigured window
will resemble that shown in Figure A-1. The reconfiguration is local, applying only to the current table, and is not
persistent.
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Figure A-1: Closest Reactions Sorted by Allele1 Match
Changing Batch Results Column Order
You can modify the Batch Results Grid table so that its columns appear in a different order.
Figure A-2: Rearranging Table Column Order
For example, as shown in Figure A-2, you can rearrange the columns so that the Patient IDs appear directly to the right
of the Sample IDs by dragging the Patient ID header from its present location and dropping it next to the Sample ID
header. You can save the modified column order to that it is persistent from session to session. See the discussion to
Figure 5-3 for details on adding or deleting columns from this table.
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Navigating the LTI Interface
Navigating Between Windows Using the Keyboard
Many of the functions and navigation features in LTI can be accessed using the hot-key functionality familiar to users
of Microsoft Windows.
To navigate between tabbed windows:
•
Select any tab in a set of tabbed windows to establish the focus on that set of tabs. Thus, to navigate in the batch
windows, select either the Batch Results, Bead Analysis, or Control Values tab.
•
Select ctrl - tab to navigate to the next tab in series
•
Select shift ctrl - tab to navigate to the previous tab in series
•
Press enter to access the window
Accessing LTI Functions via Hot Keys
After you launch LTI, press the alt key to enable the hot keys. The hot key characters for the function will be underlined on the appropriate control. You can execute the function by using the alt + key combination.
Changing Date Formats in HLATools
The date format is determined by Windows system settings.
To change the date format in Windows XP:
•
Select Start > Settings > Control Panel > Regional and Language Options.
•
In the Regional Options tabbed window, select the desired region from the pull-down list. HLATools will use the
date format displayed in the Short date text field.
To change the date format in Windows 2000:
•
Select Start > Settings > Control Panel > Regional Options.
•
In the Date tabbed window, select the desired date format from the Short date format pull-down list.
Editing the Assignment Subview Comments List
The Assignment Subview contains a pull-down list which contains frequently used stock phrases that you can add to
sample-specific comments. Any user-defined comment that you include in the sample-specific comments is timestamped and shows your username. You can edit these stock phrases using the Comments Editor utility:
To edit the stock phrases:
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•
Select Start > Programs > One Lambda > HLATools Configuration Suite> Comments.
•
Enter a new comment in the -Comments- field in the first line of the listing, or edit one of the existing comments
and OK your edits. The changes will be available when you restart HLATools.
Figure A-3: HLATools Comments Editor Tool
•
You can delete a comment by either of the following means:
•
Highlight the table row by clicking in the left-most column; press Delete from the keyboard and follow the
prompts; or
•
Select the comment text, press Delete from the keyboard, then Apply; the row will disappear from the User
Comments table.
Specifying the Data Transfer File Export Type
Some institutions may wish to employ proprietary data transfer formats when exporting results from HLATools. Unless
otherwise specified, the default export file type is compatible with Microsoft Excel. Please contact One Lambda
Customer Support to obtain the identifier to enter in the Export Type field (Figure A-4) that is unique to your institution. The export type identifier can be specified in the HLATools LABType Configuration panel. Specifying a unique
export file type can be used to invoke some or all of the following report or display behaviors:
•
Custom export file format
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•
Differing criteria for sample acceptance and review (e.g. – requiring that samples be reviewed by more than one
analyst)
•
Alternative designations for allele assignments (e.g. – computer allele assignments truncated to type and subtype
digits only)
•
Rescaling of Reaction Pattern Histogram and Bead Analysis Histograms so the entire width can be viewed without
scrolling.
•
Alternative display of analysis results in the Batch Results Grid.
Figure A-4: HLATools LABType Configuration Panel
Specifying the Character Encoding Type
Encoding is the process of transforming a set of Unicode characters into a sequence of bytes. The Unicode standard
assigns a code point (number) to each character in every supported script. A Unicode Transformation Format (UTF) is
a way to encode these code points.
HLATools supports the following options to accommodate different language formats in report generation.The ANSI
encoding type is the default as explained below. You can specify a non-default encoding type in the LABType Configuration panel (Figure A-4):
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•
ANSI – the default encoding uses the system’s current ANSI code page which is determined by the input language
specified in the Regional and Language Options control panel. A full listing of code pages can be found at
http://msdn.microsoft.com/library/default.asp?url=/library/en-us/intl/unicode_81rn.asp. You can verify which
code page your computer is using by selecting Start > Programs > One Lambda > Maintenance > Connection
Diagnostics and finding the value assigned to the CodeSet variable under the Operating System Information
section.
•
ASCII – encodes Unicode characters as single 7-bit ASCII 7-bit code representing 128 characters and control
codes. This encoding only supports character values between U+0000 (0) and U+007F (127): code page 20127.
•
BIGE – encodes Unicode characters in big-endian byte order: code page 1201
•
UTF-16 – encodes Unicode characters in little-endian byte order: code page 1200
•
UTF-8 – encodes all Unicode characters using 8-bit encoding; code page 65001
•
UTF-7 – encodes all Unicode characters using 7-bit encoding; code page 65000
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Appendix B
HLATools™ FAQ
This appendix addresses a number of issues that may arise when you are setting up or using HLATools™ LABType
Interactive.
Frequently Asked Questions
How can I tell if HLATools is using the most recent LABType products?
1. Check the product listing displayed in the HLATools Product Information table which you can access by launching
the HLA Tools Configuration Suite and then selecting HLA Update > View.
2. In your internet browser, access the One Lambda website http://download.onelambda.com/pub/tray_info/
Windows/HLATools/Labtype_Interactive/ and compare the product listing. The most recent LABType products
will appear on the download site; and they will also appear in the HLATools Product Information table
(Figure 7-2). For more on product revision numbers, see Products > Update, p. 75
Why can’t I find the Admin Diagnostics tool?
In the Version 1.0 release of HLATools, many of utility functions were carried out by standalone applications such as
the Admin Diagnostics Tool and the Quick Connect Tool. These and a number of other utilities have been combined
into the HLATools Configuration Suite which is accessible by selecting Start > Programs > One Lambda >
HLATools > Configure from the Windows desktop. For example, the functions of the Admin Diagnostics Tool and the
Quick Connect Tool are now accessed by the Configuration Suite User Management and Connections utilities,
respectively.
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Update Database – Why can’t I run …?
Users without administrative privileges will discover that the Update and Clean buttons are greyed out in the HLA
Tools Configuration Suite > Database > Local Create panel. This is because ordinary users do not have Write access
to the folder where the LTI database resides.
If you encounter this problem, ask an Administrator to log on and run these functions for you.
LTI database does not exist – Why do I get a message that …?
This message may be generated if you, as an ordinary user, have attempted to run Update Database. You are able to
start the process that stops the SQL server service and detaches the existing database. However, you cannot complete
the Update because you lack Write access to the location where the database resides. When an Administrator then
attempts to run Update Database, the application cannot find the database because you have detached it.
This problem can be solved by an Administrator using the HLATools Configuration Suite Connections tool to
reattach the database:
1. Select Start > Programs > One Lambda > HLA Tools Configuration Suite > Connections
2. Select a Connection from the Connections list (this will probably be Local)
3. Click Modify
4. The information in Sections 1 and 2 will probably not need to be changed.
5. In Section 3, select the Attach a database file as a database name option to enable the two edit fields.
6. In the first field, enter “LTI” (without the quotation marks).
7. Use the browse button to the right of the second field to locate the database file. The pathname is typically Program Files\Microsoft SQL Server\ MSSQL$ONELAMBDA\Data\LTI.mdf
8. Click Set to attach the database.
9. A user with administrator permissions can now run Update Database.
Remote LTI database – Why can’t I connect to … using Windows XP?
The Windows XP firewall on your computer may be preventing you from accessing the remote database. You may
need to open a port through your firewall.
In addition, the local security policy on the remote computer may need to be changed from its default setting. In
Windows XP, the default security setting forces all local users to access the LTI database as ‘Guest’. It is necessary to
change the security settings on the remote computer to allow other users on the network to authenticate as themselves
rather than as ‘Guest’.
You must have administrative privileges to change the firewall port settings.
Open a port though the XP firewall – How do I …?
1. From the Windows XP desktop, select Start > Settings > Control Panel > Windows Firewall > Exceptions. If
there are no entries in the Programs and Services field for tcp and udp (Figure B-1), the firewall is blocking
access to the remote database.
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2. First, add a tcp port. In the Programs and Services options, check File and Printer Sharing, then click
Add Port …
3. In the Add a Port dialog, select the TCP option and enter the following values:
Name: 1433 tcp
Port number: 1433
OK the entries. The Programs and Services field will now contain a new entry: 1433 tcp.
4. Next, add a udp port. Click Add Port … a second time.
5. In the Add a Port dialog, select the UDP option and enter the following values:
Name: 1434 udp
Port number: 1434
OK the entries. The Programs and Services field should now appear similar to Figure B-1.
Figure B-1: Modifying Windows XP Firewall Settings
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6. OK the changes. You should now be able to connect to the remote LTI database.
Local security policy on XP – How do I modify …
You must have administrative privileges to change the security settings.
1. From the Windows XP desktop, select Start > Settings > Control Panel > Administrative Tools >
Local Security Policy.
2. In the Security Settings file tree, open the Local Policies folder and select Security Options.
3. In the Policy/Security Setting listing, locate the security policy and its setting:
Network access: Sharing and security model for local accounts
Guest only - local users authenticate as Guest
4. Double-click on the name of policy to open the like-named dialog box. Select the Classic option as shown in
Figure B-2
Figure B-2: Changing the Local Security Policy in Windows XP.
5. OK the change. Other users will now be able to access the local LTI database using their own authentication.
6. Do this on all computers that will be sharing LTI database access.
Clean Database – Why do I get a BCP error …?
If you are using HLATools on an non-English language version of the Windows operation system, your MS SQL
Server Desktop Engine 2000 (MDSE) may not have been updated to the necessary Service Pack 3 product level owing
to language differences that were not recognized by the installation utility.
There are several ways to identify which version and product level of MSDE is installed on your computer:
1. From the desktop, select Start > Settings > Control Panels > Add/Remove Programs.
2. Locate the Microsoft SQL Server Desktop Engine (ONELAMBDA) entry.
3. Click on the entry to expand it, then click on the support information hyperlink. The correspondence of version
number and product level is as follows.
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Table B-1: MDSE Version and Product Level
Version
Product Level
8.00.194
RTM
8.00.384
SP1
8.00.534
SP2
8.00.760
SP3
8.00.766
SP3a
4. Alternatively, select Start > Run from the Windows desktop and execute the svrnetcn command to access
the.SQL Server Network Utility. (This requires administrative privileges.)
5. Open the Network Libraries tabbed window. The version numbers of the networking protocols are listed here
(Figure B-3).
Figure B-3: Networking Protocol Versions
If you do not have Service Pack 3 installed, update the ONELAMBDA instance of MSDE as follows:
1. Log in as administrator.
2. Access the SQL Server 2000 Service Pack 3a download page at http://www.microsoft.com/sql/downloads/2000/sp3.asp.
3. From the download drop-down list, select the language version you wish to download and press GO. This will take
you to the download page for the specific language. At the bottom of this page are several download links.
4. Choose the file of the format
<lang>_sql2kdesksp3.exe
where <lang> is the abbreviation for the language you have selected.
5. Download this file to your desktop.
6. Click on the file. This will open an Installation Folder dialog. Accept the suggestion for the default installation
folder (C:\sql2ksp3) unless you want to save the file to a different location. The file will be unpacked in this folder.
7. From the desktop, select Start > Programs > Accessories > Command Prompt.
8. Navigate to the installation folder mentioned above.
9. Execute the command:
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setup /upgradesp sqlrun INSTANCENAME=ONELAMBDA DISABLENETWORKPROTOCOLS=1
To update the default instance of MSDE for other applications that may use it other than LTI:
1. Repeat steps 7 through 8 above.
2. Execute the command:
setup /upgradesp sqlrun DISABLENETWORKPROTOCOLS=1
For more information on SP3/SP3a, see article 819334 on the Microsoft Support website.
Why do I have to log into the database when I try to print some reports?
If a Database Login dialog appears when you try to generate long reports such as the Patient Summary Report by
Batch, it means that the TCP/IP and Named Pipes protocols have not been enabled in your Server Network Settings.
You can ignore the request for login. However, the time required to print out the requested report may be inconveniently long.
The steps required to enable these protocols so that all reports are generated and printed quickly are described in detail
in Peer-to-Peer Installation, p. 135.
Windows Authentication Mode – Why can’t I set up …?
If your computer uses the Windows 2000 operating system and you have chosen Windows Authentication as your
security mode as described in HLATools Security Modes, p. 149, you may find that you are unable to log in to LTI. This
is due to the fact that Version 8.0 of the Microsoft SQL Server Desktop Engine (MSDE) does not support the Windows
authentication mode on systems running Windows 2000. This problem will be rectified with an upcoming release of
the MSDE software.
Alleles – How are they designated?
The HLA locus (A, B, C, DR, DP or DQ) is designated first, followed by an asterisk. The allele type and subtype are
designated by four digits. The first two digits designate the serological equivalent of the antigen. The second pair of
digits designates the specific allele. Additional digits may be added that indicate a separate, higher resolution subtype
that is produced by a synonymous nucleotide substitution (one that does not change the encoded amino acid, and thus
does not affect the antigenic expression of the protein on the cell surface) or an intron polymorphism (a mutation that
occurs outside the region that encodes the amino acids in the protein on the cell surface).
Table B-2: HLA Allele Nomenclature
Locus
*
1
2
3
4
DRB1
*
11
DRB1
*
11
01
DRB1
*
11
01
01
01
DRB1
*
11
01
01
02
5
Resolution
Low
High
High
N
High
Where:
1. Allele type (equivalent of serological antigen)
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2. Allelic subtype (amino acid difference)
3. Non-coding polymorphism (e.g. synonymous nucleotide)
4. Intron polymorphism
5. Expression level of the allele
•
A – indicates 'Aberrant' expression where there is some doubt as to whether a protein is expressed
•
C – indicates an allele product that is present in the 'Cytoplasm' but not on the cell surface
•
L – has been shown to have 'Low' cell surface expression when compared to normal levels
•
N – has been shown not to be expressed
•
Q – indicates 'Questionable' expression in that the mutation seen in the allele has previously been shown to
effect normal expression levels, but the expression remains unconfirmed
•
S – indicates that the allele codes for a protein that is expressed as a soluble 'Secreted' molecule but is not
present on the cell surface
As of December 2005 (per the Anthony Nolan website), no alleles have been named with the 'A' or 'C' suffixes.
Table B-3: HLA Allele Nomenclature Examples
Nomenclature
Meaning
DRB1
Locus
DRB1*11
Group of alleles that encode the DRB1*11 antigen
DRB1*1101
Specific allele
DRB1*1160N
Specific null allele
DRB1*110101
Allele that differs by a synonymous polymorphism
DRB1*110102
Allele that differs by a synonymous polymorphism
DRB1*110103
Allele that differs by a synonymous polymorphism
DRB1*11010101
Allele containing a variation in a noncoding region (intron polymorphism)
DRB1*11010102N
Null allele containing a variation in a noncoding region (intron polymorphism)
DRB1*1101-03
Ambiguity: DRB1*1101 or DRB1*1102 or DRB1*1103
DRB1*11MN
The above using NMDP ambiguity code
DRB1*1101/03
Ambiguity: DRB1*1101 or DRB1*1103
DRB1*11AC
The above using NMDP ambiguity code
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Matched Reaction Pairs (MRPs) – How does LTI identify …?
Figure B-4 illustrates how LTI determines Matched Reaction Pairs (pairs of alleles whose combined reaction patterns
exactly match the observed sample reaction pattern), Close Reactions (pairs of alleles whose combined reaction
patterns differ by one bead reaction - positive or negative - from matching the observed sample reaction pattern) and
False Reactions (pairs of alleles whose combined reaction patterns differ by one or more bead reaction - positive or
negative - from matching the observed sample reaction pattern).
Before starting any of these procedures LTI compares the normalized mean fluorescence values for each of the beads in
the sample to the established cutoff values for the beads to determine which beads have registered positive reactions.
Based on this comparison LTI constructs the Sample Reaction Pattern for the current sample (see Figure 5-8, Assignments View with two Matched Reaction Pairs). The reaction patterns for all samples in a batch are determined when the
initial batch import in done.
1. Matched Reaction Pairs – LTI makes an exhaustive comparison of the combined reaction patterns of all pairs of
alleles recognized by the selected LABType product to the observed Sample Reaction Pattern. The two alleles in
each comparison are designated Allele I and Allele II.
•
LTI steps through the entire list of alleles, combining the reaction pattern of the first Allele I with each of the
other alleles (each in turn considered as Allele II) and comparing the combined reaction patterns to the Sample
Reaction Pattern (SRP). If any combined pair matches the SRP, that pair is recorded as a Matched Reaction
Pattern (MRP). (This may be thought of as an inner comparison loop).
•
LTI then treats the next allele as Allele I and repeats the combining/comparing procedure with all of the other
alleles. (This may be thought of as an outer comparison loop). Since any given allele a plus allele b is the same
as allele b plus allele a, the combining/comparing is only done for the first case. As a result, the total number of
combining/comparing steps is (n x n)/2 where n is the number of alleles recognized by the LABType product.
The combining/comparing procedure is executed when the user accesses the Assignments View for a given sample
either by clicking on the sample in the Batch/Sample file tree (Figure 5-2) or on the sample row in the Batch
Results Table (Figure 5-3).
2. Close Reactions – LTI identifies Close Reactions by toggling the bead reactions in the Sample Reaction Pattern
one by one (from positive to negative or negative to positive). For each change in bead state, LTI repeats the combining/comparing procedure described above, looking for allele pairs whose combined reaction patterns match that
of the now-modified Sample Reaction Pattern.
The search for Close Reactions is executed when the user accesses the Close Reactions subpanel (Figure 5-13) in
the main Assignments View or the Bead Analysis View (Figure 5-9). For each Close Reaction detected, LTI reports
the allele pair and ID of the involved bead in the Closest Reactions subpanel (Figure 5-13).
3. False Reactions – LTI searches for False Reactions by looking at all allele pairs with combined reaction patterns
that are one bead reaction removed from yielding a Matched Reaction Pair, or a “one-away”. (A “one-away” False
Reaction is also a Close Reaction.) LTI then searches for “two-aways”, “three-aways”, and so forth. Searches for
False Reactions are initiated when the user changes values for bead cutoffs, either globally in the Bead Analysis
Panel (Figure 5-4) or locally from within a sample’s Reaction Pattern Histogram (Figure 5-9).
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Figure B-4: LTI Analysis Flowchart
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MRP Table – How does LTI populate the …?
The process that LTI employs to populate the MRP table can be illustrated by referring to the Reaction Grid which is
discussed in some detail in Chapter 5 (see Reaction Grid, p. 58).
•
The x-axis lists the beads in the assay, including the control bead(s). In the default view, the beads appear in ascending order from left to right (Figure B-5). (These are the bead numbers in the assays, not the proprietary OLI
bead identifiers.) Typically there are about 100 different kinds of probes in an assay and several hundred beads with
each type of probe. When the reaction grid for a sample is first accessed, the topmost row of the table contains the
sample reaction (Rxn). The beads with positive reactions are indicated by columns with light blue background.
Clicking on a row header cell causes all the reactions for that allele to be clustered to the left. In Figure B-6 this has
been done for the sample reaction itself.
Figure B-5: Reaction Grid – Default View
•
The y-axis lists all the alleles in the assay. Many of the entries in the Allele column are single, unambiguous alleles
which have wholly numerical designations. Other entries designate groups of alleles with shared specificities and
are designated with alphanumeric codes.
•
When multiple alleles have a common reaction pattern, the ambiguity is indicated using an NMDP or local ambiguity code. For example, in Figure B-6, the alleles DRB1*010101, DRB1*010102, DRB1*0107, DRB1*0107,
DRB1*0108 and DRB1*0111 all share a common reaction pattern. This group of alleles is designated in the Allele
column by the NMDP code DRB1*01EW. The member alleles designated by an ambiguity code are displayed in
the table header space when you click anywhere in an allele’s row.
•
LTI compares the observed bead reactions with specificities represented by the alleles in the Allele column and
marks the positive reactions with a red x.
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Figure B-6: Reaction Grid – Golden Alleles
•
Alleles (or groups of alleles) with reaction patterns that are subsets of the sample reaction pattern are termed “golden” alleles, and are highlighted by a light yellow background (dotted outline in Figure B-6). If the intersection of
two golden alleles completely matches the sample reaction pattern, the pair is termed a “Matched Reaction Pair”.
In the sample shown in Figure B-5 and Figure B-6 the intersections DRB1*01EW/DRB1*14WHG and
DRB1*010103/DRB1*14WHG both yield MRPs (Figure B-7).
Figure B-7: Matched Reaction Pairs
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MRP Table – How are Allele1 and Allele2 grouped in the …?
When LTI analyzes the observed sample reaction pattern, it creates an exhaustive listing of all known alleles that can be
grouped into pairs consistent with the observed Sample Reaction Pattern (SRP). These pairs are the Matched Reaction
Pairs (MRPs) shown in Figure B-8. However, before the MRPs can be shown in the arrangement seen in Figure B-8,
the alleles making up the MRPs need to be systematically sorted between Allele1 and Allele2. To illustrate the sorting
method, we will consider a simple case in which all of the alleles have the first two digits in common. The same
method is applicable when the first two digits are different. For the sake of simplicity, only the first four digits are
considered in this sorting routine.
Table B-4: Grouping Alleles for the MRP Tabulation
Original Pairing
Simplified Final Pairing
Row
Allele 1
Allele 2
Allele 1
Allele 2
1
0101
0101
0101
0101
2
0101
0102
0102
3
0101
0103
0103
4
0101
0104
0104
5
0101
0105
0105
6
0101
0106
7
0102
0103
0103
8
0104
0105
0105
9
0106
0106
0106
0106
Three rules are observed:
1. Trivial duplicate entries in a column are avoided. Thus, in rows 2 through 6, 0101 is not repeated in Allele 1, but
0102, 0103, etc., are displayed in Allele 2 because each instance is a first occurrence.
2. The entries for the columns are swapped if one or the other column already contains the entry. Thus, in row 7,
Allele 2 already contains an instance of 0103, forcing the entries to be swapped. 0103 is now grouped under
Allele 1, while 0102 is deleted under Allele 2 because there is already an instance of 0102 in that column. The
same holds true for the entries in row 8.
3. A homozygous allele is represented in both columns (row 9).
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In the MRP tabulation and elsewhere, high resolution alleles are designated with four digits: the first pair designates the
allele family (equivalent of the serological antigen) and the second pair the allelic subtype (amino acid difference).
Examples would be B*4435 or B*5707 in Figure B-8. Groups of alleles with shared reaction patterns are designated
with two digits and an NMDP or local ambiguity code. For example, B*44ANYV signifies that the allele may be
B*440301, B*4426, B*4436, B*4438 or B*4439. Further digits or characters that are used to denote higher resolution
alleles (noncoding or synonymous polymorphisms, intron digits and null or low levels of expression) may also be
shown in the Matched Reaction Pairs tabulation. See the FAQ Alleles – How are they designated?, p. 116.
Figure B-8: Matched Reaction Pair Tabulation
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All Alleles Table – How is the … generated?
The All Alleles (Recognized Allele Pairs/Serological Equivalent) table is created by matching all possible combinations of the alleles in the Allele1 cell of the MRP table with all possible alleles in the Allele2 column cell.
For example, in the first row of the MRP table, B*44ANYV indicates that allele1 may be B*440301, B*4426, B*4436,
B*4438 or B*4439 (5 possibilities) and the matching allele2 B*57XX4 may be B*570301 or B*570302
(2 possibilities). Together, these yield 10 pairs as indicated by the cells in Figure B-9 marked with rectangles. A
complete matching of the possibilities from the cells in both columns yields the twenty pairs contained in the table of
Figure B-9.
Figure B-9: Recognized Allele Pairs/Serological Equivalents
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Type/Subtype Allele Assignments – How are … made?
This FAQ addresses how the Type/Subtype table is generated from the entries in the All Alleles table, Table B-4.
1. All digits beyond the first four are dropped (columns 1 and 2 in Table B-5).
2. All resulting duplicate pairs are discarded (columns 3 and 4).
3. All duplicate entries in each column are discarded (columns 5 and 6). The resultant columns are identical to those
shown in the Type/Subtype listing (Figure B-10).
Table B-5: Creating the Type/Subtype Tabulation
Truncated to 4 Digits
Allele 1
Duplicates Pairs Discarded
Duplicate Entries Discarded
B*44AXJR
B*57CG
Allele 2
Allele 1
Allele 2
Allele 1
Allele 2
B*4403
B*5703
B*4403
B*5703
B*4403
B*5703
B*4403
B*5703
B*4403
B*5707
B*4403
B*5707
B*4426
B*5703
B*4426
B*5703
B*4426
B*5703
B*4426
B*5707
B*4426
B*5707
B*4430
B*5707
B*4430
B*5707
B*4430
B*4435
B*5703
B*4435
B*5703
B*4435
B*4435
B*5703
B*4435
B*5707
B*4435
B*5707
B*4436
B*5703
B*4436
B*5703
B*4436
B*5703
B*4436
B*5707
B*4436
B*5707
B*4437
B*5707
B*4437
B*5707
B*4437
B*4438
B*5703
B*4438
B*5703
B*4438
B*4438
B*5703
B*4438
B*5707
B*4438
B*5707
B*4439
B*5703
B*4439
B*5703
B*4439
B*5703
B*4439
B*5707
B*4439
B*5707
B*4426
B*5707
B*4436
B*4439
As a final step, LTI assigns NMDP codes to all alleles that share the same reaction pattern. If the assignment is
unambiguous (i.e. the candidate alleles contain no false reactions and are of the same type), the alleles in each table are
again represented by a more inclusive NMDP code and this assignment is displayed in the column header. If a single
code assignment cannot be made because the alleles are of different types or they contain a false reaction, the typing is
considered ambiguous and is denoted by an orange background in the Type/Subtype subview.
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Figure B-10: Type/Subtype Tables
Reaction Grid – How is the … sorted?
The following heuristic method is employed to sort alleles in the Reaction Gird to allow the user to examine how
similar they are to the Sample Reaction Pattern. The allele sorting is driven by two user-specified alleles (referred to
below as “First Allele” and “Second Allele”) which are entered in the text search fields at the top of the grid
(Figure 5-16). Five different types of scores are assigned to each allele: A1, A2, PP, FN and TP. Typically these scores
are not displayed in the Reaction Grid.
1. A1 scores are assigned to the sample reaction and to the alleles as follows:
•
The Sample Reaction is always scored “1”.
•
Alleles that share fewer than 75% positives with the First Allele are scored “0”.
•
Alleles that share more than 75% positives with the First Allele are scored “5”, unless
•
they also share the first four digits (family and subtype designations) with the First Allele, in which case they
are scored “3”. (There will always be at least one “3”.) If an allele designation includes an NMDP or local ambiguity code, the comparison is made on the basis of the numeric allele designations defined by the alphabetic
code.
2. A2 scores are assigned to the sample reaction and to the remaining alleles after the positive reactions have been
clustered to the left of the grid. The “left” part of the gird is defined as the string of consecutive positives in
Allele 1 when its positives are pulled to the left. The rest of the grid is defined as the “right” or the “Allele 2
remainder”. Two scoring cases are distinguished.
A. No specification is made in the Second Allele entry field.
•
The sample reaction is always scored “5”.
•
Alleles that share more than 75% positives with the Second Allele are scored “5”.
•
Alleles that share fewer than 75% positives with the Second Allele are scored “0”
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B. A specification is made in the Second Allele entry field.
•
The sample reaction is always scored “5”.
•
If the type or subtype designation (first two and fours digits, respectively) of the allele matches that of the
Second Allele and the allele shares more than 75% positives, it is scored “5”.
•
If the type or subtype designation (first two and fours digits, respectively) of the allele does not match that
of the Second Allele, but the allele shares more than 75% positives, it is scored “3”.
•
Alleles that share fewer than 75% positives with the Second Allele are scored “0”.
3. PP scores can be assigned after the positive reactions have been clustered to the left of the grid.
•
The sample reaction is always scored as –(TPs in Sample Reaction – TPs in First Allele). This is always a negative number.
•
For alleles with a positive A1 score, PP is the number of true positives shared with the First Allele. This is always a positive number.
•
For A2 alleles, i.e. with a “0” A1 score, PP is the negative of the number of positives on the right side of the
positives field in common with the sample reaction.
•
When the change in PP from the initial A2 allele PP score is greater than 2, a score of “0” is assigned.
4. FN scores are:
•
For each allele, the number of False Negatives plus 1.
•
By definition the Sample Reaction is “0”.
5. TP scores are:
•
For each allele, the number of True Positives.
•
The Sample Reaction is assigned a TP score of “101”, which is larger than any possible number of True Positives. (No assay has more than 100 different types of beads.) This assures that the Sample Reaction always subsorts as the first item in the “0” set of False Negatives.
6. The alleles are then sorted and subsorted in this order:
•
A1 descending
•
A2 descending
•
PP ascending
•
FN ascending
•
TP descending
To illustrate the sorting protocol, Table B-6 reproduces part of a reaction grid with alleles scores for A1, A2, PP, FN
and TP. Deleted columns and rows are indicated by the tilde sign.
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Table B-6: Allele Sorting in the Reaction Grid
Allele
11 31 52 54 64 43 47 56 57 72 75 20 21 23 38 39 46 60 69 77 58 63 66
A*0302
x
x
x
x
x
x
A*0305
x
x
x
x
x
x
A*0307
x
x
x
x
x
x
x
A*0303N
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
~
x
x
x
x
A1 A2 PP FN TP
5
0
10
2
11
5
0
11
1
12
5
0
11
1
12
5
0
11
1
11
5
0
11
1
11
5
0
11
2
11
5
0
11
2
11
5
0
11
2
11
A*0306
x
A*0308
x
A*0309
x
x
x
x
x
x
x
x
x
x
A*0312
x
x
x
x
x
x
x
x
x
x
x
A*030102
x
x
x
x
x
x
x
x
x
x
x
3
0
11
1
11
A*03ANGK
x
x
x
x
x
x
x
x
x
x
x
x
3
0
12
1
12
Rxn
x
x
x
x
x
x
x
x
x
x
x
x
A*68NPN
x
x
x
x
x
A*6815
x
x
x
A*6827
x
x
x
x
x
A*3403
x
x
x
x
x
A*6823
x
x
x
x
x
~
~
~
~
~
~
x
~
~
x
~
~
~
x
x
x
~
x
x
x
x
x
x
x
x
1
5
-7
0
101
x
x
x
x
x
x
x
0
5
-7
1
12
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
~
~
~
~
x
~
x
x
x
x
x
~
~
~
~
~
~
~
~
0
0
-7
3
10
0
0
-6
2
11
0
0
0
2
10
0
0
0
2
10
~
~
~
~
~
NMDP Codes – How can I automatically update …?
The easiest way to assure that HLATools is using the most current NMDP codes is to update them automatically.
Windows XP and Windows 2000 both include a utility that you can use to set up an automated task or “chron job” to
update the codes.
In an ordinary installation of HLATools, the NMDP updating utility InputNumericTxt.exe is located in C:\Program
Files\One Lambda\HLATools. You can use the Windows Scheduled Tasks utility to run this executable automatically. The following steps describe the setup in Windows XP. Some terms may be slightly different in Windows 2000.
1. From the Windows desktop, select Start > Settings > Control Panel >Scheduled Tasks.
2. Launch the Add Scheduled Tasks application.
3. The Scheduled Task Wizard opening dialog will appear. Click Next to proceed.
4. Locate and select Update NMDP Data in the applications listing, or you can browse to locate the utility. Click
Next to proceed.
5. The name of the selected application will automatically populate the task name text entry field. You can rename the
task if you want to.
6. Specify a Start Time and Start Date. For performance frequency, select Weekdays, since the codes are updated
daily Monday through Friday by NMDP Research. Click Next to proceed.
7. An authorization dialog appears next. Your username (or that of the current user if someone other than you is
logged on to the computer) appears in the first text field. Enter and confirm your Windows log-on password. Click
Next to proceed.
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8. If the password matches the username, Windows will schedule the automatic task as shown in Figure B-11. This
listing also lists when the last time the utility was run and when it is next scheduled to run. Click Finish to complete the automatic scheduling. You can modify schedule settings by double-clicking on the scheduled task in the
listing.
Figure B-11: Schedule Tasks – Updating NMDP Codes Automatically
LTI database directory – Why is the … growing excessively?
Your LTI database directory may grow to excessive size if you run Update Database frequently without purging your
system of backup database files. How this happens becomes clear when we review the steps executed by the HLATools
Update Database utility.
1. To start, the Update Database script creates a new folder called C:\Program Files\Microsoft SQL
Server\MSSQL$ONELAMBDA\data\backupYYYYMMDDhhmm and saves copies of your current .mdf and
.ldf database files to this folder. These copies are called simply <dbName>.mdf and <dbName>.ldf. (The default
<dbName> is LTI.) This backup copy of the .mdf database file is detached from your MSDE SQL server instance
to assure that all of its data remains secure. (The .mdf file is the primary data file and the .ldf file is a log file.)
2. Next, the script renames the current <dbName>.mdf and <dbName>.ldf database files, calling them
temp<dbName>YYYYMMDDhhmm.mdf. These copies reside in the ~\data folder. The
temp<dbName>YYYYMMDDhhmm.mdf is reattached to your MSDE SQL server instance.
3. The script runs Clean Database, creating a new database called <dbName>.mdf in the ~\data folder, and merges
the data in the renamed temp copy with the new <dbName>.mdf database. At this point the combined size of your
database structure will be about three times the original size, consisting of the new database, the backup and the
temporary copy of your original. Thus, if you run Update Database frequently, the proliferation of backup and
temporary files may consume a great deal of disk space.
4. To avoid this and to protect your valuable data, we recommend that you save the backup copies of your databases
by burning them onto a CD or other storage medium before deleting them from your system.
5. Before you copy or delete a database, you must detach it from your MSDE SQL server instance: select Start >
Programs > One Lambda > HLATools Configuration Suite > Database > Attach/Detach from the Windows
desktop.
6. Select the Detach Database option.
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7. Select the attached database from the pull-down list and click Detach.
Figure B-12: Attach/Detach Database Tool
How do I update an LTI database with a non-default name?
The name for the LTI database is set by assigning a value to the TABLE variable in the ALL.bat batch file. The default
name for LTI databases is “LTI”. However, in the example shown in Figure B-13, the LTI database is called “HUGO”
instead of “LTI”.
You can update a database with a non-default name by running the batch file updateDB.bat which is located in the
C:\Program Files\One Lambda\HLATools\Clean Database\UpdateDB\ folder. Before running updateDB.bat, you
must be sure that the database file name there is identical to the name in ALL.bat (Figure B-14). (ALL.bat is located
in the ~\Clean Database\ folder.) All.bat creates a new, clean database.
The steps which the system executes when updating a database are briefly as follows. The system…
1. Creates a folder with the name Backup and a timestamp.
2. Detaches the database.
3. Copies the databases .mdf (main data file) and .ldf log file into the newly created backup directory.
4. Renames the original .mdf and .ldf files by prefixing the word temp and appending the timestamp to the original
filename.
5. Attaches the newly renamed files.
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6. Runs the All.bat file, creating a new database which contains the latest LABType product information, NMDP and
serological data.
7. Runs MERGE, which merges the temp copy of the database with the newly created database described in the previous step.
For detailed names of the files and folders involved in the updating process, see the FAQ, “Why is the LTI database
directory growing excessively?”
Figure B-13: Setting Database Name in ALL.bat
Figure B-14: Setting Database Name in updateDB.bat
How do I install MSDE to a non-default folder?
You must have administrative permissions to perform this installation.
To Install MSDE to a Directory other than Default
1. Go to: http://msdn.microsoft.com/sql/200/downloads/default.aspx
2. Click the MSDE 2000 Release A link.
3. Select the language version you would like to download and click Change.
4. Scroll down the page and click Download for the <lng>_MSDE2000a.exe file,
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where <lng> is the 3-letter abbreviation for the language you have selected, i.e. GER_MSDE2000a.exe for
German or JPN_MSDE2000a.exe for Japanese.
5. Double-click on the file and select a directory to extract the installation files into.
6. Select Start > Programs > Accessories > Command Prompt. Using the command prompt, navigate to the extraction directory.
To install to a default instance, enter:
setup SAPWD="AStrongPassword" TARGETDIR="C:\MyInstanceFolder\"
To install to a named instance, enter:
setup SAPWD="AStrongPassword" INSTANCENAME="ONELAMBDA"
TARGETDIR="C:\MyInstanceFolder\"
Note that the directory specified for the “TARGETDIR” property must always include a trailing back-slash.
How can I import LABType Classic analyses into HLATools?
The LABType Database Upgrade utility allows you to import analysis results from a LABType FoxPro database into an
SQL database. When you migrate a LABType FoxPro database, the installation program checks whether the required
Microsoft .Net and MSDE software components are present on your system and will install them, if necessary. In
addition, the installation program will create an SQL database if none is present. These actions are executed automatically and do not require user intervention.
To migrate your data from a LABType FoxPro database into an SQL database:
1. Locate the Database Upgrade folder on your LABType distribution CD and launch the setup.exe inside.
2. Follow the InstallShield Wizard instructions, accepting the usage agreement terms and the default values. Just
before finishing the installation, the InstallShield Wizard creates an Upgrade LABType Database icon on your
desktop. You can launch the Upgrade LABType Database application in either of the following ways:
•
Select the Launch LABType Import checkbox, then Finish, or
•
Exit the installation by clicking Finish, then launch the application using the icon.
3. If LABType is installed in the default C:\LABType directory, you can proceed directly to the upgrade step. If not, a
directory browse window will appear. Browse to locate the directory containing the LABType executable
labtypeN.exe, where N is the version number (Figure B-15).
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Figure B-15: Locating the LABType Classic Folder
4. The installation program will check to see if the ONELAMBDA MSSQL server instance has the proper SQL
Server network protocols enabled. If not, the program will enable it. This occurs without user intervention.
5. The installation program next checks to see if an SQL database exists on the server instance. If an SQL database
does exist, the program makes a backup copy. If one does not exist, the program will create a new database. This
may require several minutes.
6. At this point a window similar to that shown in Figure B-16 appears which lists the file paths for the LABType session files. Click Upgrade to continue.
Figure B-16: Upgrade LABType Database Files
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7. The utility performs the following steps for the samples in each batch:
A. Imports raw data (count and trimmed mean).
B. Normalizes the data.
C. Copies the LABType analysis results into the SQL database and maintains the “Locked” status of samples
whose allele assignments have been reviewed and accepted.
D. Carries over any user adjustments in bead cutoff values and notes that the cutoffs have been changed from the
default values.
E. Carries over sample-specific comments.
F. Checks to see if the Patient ID associated with the sample being imported already exists in the SQL database.
•
•
If the Patient ID exists, only information from non-blank fields is imported: i.e. no existing data is overwritten by null values. Imported patient demographic data includes:
Patient ID
Last Name
Middle Name
First Name
Race
Family Name
DOB
Relationship
ABO
Rh
If the Patient ID does not exist, the program uses the Sample ID as a substitute Patient ID and imports the
patient demographic data for the sample into the SQL database.
The steps outlined in §7 are repeated for each session file shown in Figure B-16.
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Appendix C
Network Installation of HLATools
This chapter addresses two configurations for installing HLATools™ software in a networked environment:
•
Peer-to-peer configuration where several workstations are connected in such a way that one user can access the
files and database(s) of other users.
•
Network configuration where a number of client workstations are connected to a central server. The server handles
many of the computational and databases functions for the client machines.
The standard installation of HLATools on a standalone workstation is discussed above in Chapter 2, Installation and
Log On.
Important Note: Peer-to-peer and network configuration is wholly the responsibility of the user and the user’s IT
department. HLATools is not designed to be an automated network configuration tool. For wireless network
installation, the user must ensure electromagnetic compatibility and is responsible for electromagnetic interference.
Peer-to-Peer Installation
In order to use LTI in a peer-to-peer or network configuration, it is necessary to enable two communications protocols
on each computer: Named Pipes (a protocol for exchanging information between two applications, possibly on
different computers) and TCP/IP. Enabling these protocols requires administrative user privileges.
To enable Named Pipes and TCP/IP:
1. From the Windows XP or Windows 2000 desktop, select Start > Run.
2. Enter svrnetcn in the Open field and click OK to access the SQL Server Network Utility.
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Note: If you have applications on your system that use versions of the SQL Server Network Utility earlier than 8.0,
executing the Run > svrnetcn command may launch the older utility. If this occurs, navigate to the newer
executable which can be found in C:\Program Files\Microsoft SQL Server\80\Tools\Binn and run svrnetcn.exe
from that location.
Figure C-1: SQL Server Network Utility
3. Select the ONELAMBDA instance of the SQL server installation on the local computer (in this case, swordfish).
4. Make sure that both Named Pipes and TCP/IP have been moved to the Enabled protocols field. OK the newly
enabled protocols. Any changes that have been made are thus saved, but will not go into effect until the computer
has been restarted.
5. Repeat this procedure on every computer that will be sharing LTI databases.
Creating a Database on a Remote Server
The all.cmd script is located in the C:\Program Files\One Lambda\HLATools\Clean Database folder. This script is
launched when you run the Clean Database Utility which creates an LTI database on your local computer (see Running
the Clean Database Utility, p. 18). You can use this same script to create an LTI database on a remote computer. Of
course, you must have write access to the remote computer to do this.
To create a database on a remote computer, you will need to edit the server name, the server instance name, and the
database name in the all.cmd script. These changes involve only the first few lines of the file.
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When you open all.cmd in a text editor, the pertinent lines will appear as follows (without the line numbers which are
inserted here for reference only):
1)
2)
3)
4)
5)
6)
7)
8)
9)
10)
11)
12)
@echo off
SET SERVER=(local)\OneLambda
SET TABLE=LTI
SET LOGIN=-E
REM SET LOGIN=-Usa -Psecret
REM SET SERVER=192.168.1.200\OneLambda
net start mssql$onelambda
osql -S %SERVER% %LOGIN% -q "EXIT(DROP DATABASE LTI)"
net stop mssql$onelambda
Del "%ProgramFiles%\Microsoft SQL Server\MSSQL$ONELAMBDA\Data\LTI*.*"
net start mssql$onelambda
osql -S %SERVER% %LOGIN% -q "EXIT(CREATE DATABASE LTI)"
1. In line 2, (local)\OneLambda defines the server name and server instance. Note that a server instance does not
need to be defined, in which case the database will be created in the default server instance. Alternatively, you can
specify an existing instance.
•
If you intend to run all.cmd locally, i.e. from the same server in which you are creating the new LTI database,
leave (local) unchanged. The database administrator may want the LTI database to reside on the default
server instance of the server to reduce resource usage. The default may either be unnamed (i.e. “null”) or
named. In this case, delete \OneLambda and replace it with the default (either nothing or with the name of the
default server instance). Otherwise, the script will create a separate \OneLambda server instance.
•
If you intend to run all.cmd remotely, i.e. from a different server than the one on which you are creating the
new LTI database, replace (local) with the IP address of the target server or the server name. Again, consult
your database administrator to determine if you should use the default server instance (i.e., either “nothing” or
a named instance).
2. In line 3, you can use the existing LTI database name, or change it to any name you wish, such as MY_LTI_DB.
3)
SET TABLE=MY_LTI_DB
3. Lines 4 and 5 set the login protocol.
•
The -E switch invokes Windows authentication. If you want to use Windows authentication, leave lines 4 and
5 unchanged.
•
If you want to use SQL authentication, comment out line 4 and un-comment line 5:
4)
5)
REM SET LOGIN=-E
SET LOGIN=-Usa -Psecret
In this case, the user name would be sa and the password secret. You can change both of these values as you
wish. For example, for the username/password combination myname/MyWord, lines 4 and 5 would be
4)
5)
REM SET LOGIN=-E
SET LOGIN=-Umyname -PMyWord
Usernames and passwords follow the usual conventions, with the username being case insensitive and the
password being case sensitive.
4. In line 8, replace the hard-coded value LTI with the environment variable %TABLE%:
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8)
osql -S %SERVER% %LOGIN% -q "EXIT(DROP DATABASE %TABLE%)"
5. Comment out line 10:
10)
REM Del "%ProgramFiles%\Microsoft SQL Server\MSSQL$ONELAMBDA\Data\LTI*.*"
6. In line 12, replace the hard-coded value LTI with the environment variable %TABLE%:
12)
osql -S %SERVER% %LOGIN% -q "EXIT(CREATE DATABASE %TABLE%)"
7. To create a database named MY_LTI_DB on a remote server with the IP address 199.199.1.200 using the default
“null” server instance, lines 1 – 12 should appear as below. Login would employ the SQL authentication protocol
with the username/password combination myname/MyWord.
1)
2)
3)
4)
5)
6)
7)
8)
9)
10)
11)
12)
@echo off
SET SERVER=199.199.1.200
SET TABLE=MY_LTI_DB
REM SET LOGIN=-E
SET LOGIN=-Umyname -PMyWord
REM SET SERVER=192.168.1.200\OneLambda
net start mssql$onelambda
osql -S %SERVER% %LOGIN% -q "EXIT(DROP DATABASE %TABLE%)"
net stop mssql$onelambda
REM Del "%ProgramFiles%\Microsoft SQL Server\MSSQL$ONELAMBDA\Data\LTI*.*"
net start mssql$onelambda
osql -S %SERVER% %LOGIN% -q "EXIT(CREATE DATABASE %TABLE%)"
Connecting to a Remote LTI Database
LTI is often installed in a peer-to-peer configuration where one user wishes to access LTI databases on another’s
computer. A major issue in peer-to-peer installations is the proper setting of permissions so that only authorized users
can access remote LTI databases. To allow a remote user to access a target user’s local LTI database, the target user
must add the remote user to the list of authorized users on the target computer. (This can be done remotely as well, as
will be discussed in Editing the Authorized Users List from a Remote Server, p. 146).
In setting up peer-to-peer access, it is necessary to know the network identities (full computer names) of the computers.
You can find this by selecting My Computer > Properties > Network Identification from the Windows 2000 desktop
or My Computer > Properties > System Properties > Computer Name from the Windows XP desktop. In this
example we establish connections between two servers named swordfish and skipjack.
1. On the target computer (swordfish), launch the User Management tool by selecting Start > Programs >
One Lambda > HLATools Configuration Suite> User Management.
2. Select List Users to refresh the list of users who currently have access to the swordfish LTI database (Figure C-2).
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Figure C-2: User Management Tool
3. Two methods of authentication are available to control access to the local LTI database.
A. SQL Users – remote users are granted access by means of a username/password combination different from
that used for Windows network security. See Linking to another Server using SQL Authentication, p. 147.
B. Windows Users – this option employs standard Windows network security. Anyone on your Windows network can be granted access to a target computer’s LTI database after he or she has been added to that computer’s authorized users list.
In this example we will establish permissions for a remote user named Quimby. Click Add Windows User/Group
to access the like-named dialog (Figure C-4).
Figure C-3: Simple Network Installation with two Users
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Figure C-4: Users/Groups on Windows Network
4. Make sure the Domain tab is active. The Existing Users list should already be populated. You can refresh the list at
any time by pressing List Users to load the users on the network (in this case network1) into the display field.
5. Select the name of the remote user who will be accessing this local database (in this case Quimby). Then press
Add. Quimby is now added to the list (Figure C-5).
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Figure C-5: Newly Added User with Access to Local Database
6. The LTI database on swordfish can now be accessed by two default local administrators (sa and
BUILTIN\Administrators), three local users (workshop1, tissuelab, and Homer) and one remote user, Quimby.
Now that Quimby has been added to the authorized users list on Homer’s computer (swordfish), log into
Quimby’s computer (skipjack) and launch the Connection utility by selecting Start > Programs >
One Lambda > HLATools Configuration Suite > Connections. (Later we will see how to do this remotely.)
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Figure C-6: HLATools Configuration Suite Connections Tool
7. Click Add to create a new connection (Figure C-6).
8. In the Database Settings dialog that appears (Figure C-7), enter a name for the new connection. This name can be
an alias for the target computer to which you are establishing the connection or any name you wish. Here we will
create a connection named Homer.
9. Click Set to OK the newly added name. The new name will now appear below the Local connection in the Connection Name list (Figure C-6).
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Figure C-7: Database Settings
10. The name of the server to which LTI currently has access will appear in the topmost editable field (1). Note that the
name of the newly created connection (Homer) appears in the Connection Name field (Figure C-7). The “More”
button (… accesses a list of other accessible servers in the pull-down list. If this list does not populate, check that
the Named Pipes and TCP/IP communications protocols have been enabled. (For details, see Peer-to-Peer Installation, p. 135.)
11. Accept the default options for Windows NT Integrated Security (2) and LTI for database selection in (3). Select the
desired server from the pull-down list (Figure C-8), or enter its name directly into the field. The name must observe
normal pathname naming conventions and not contain control characters or whitespace. In this example we will
select the server swordfish where we just added the remote user Quimby to the authorized users list.
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Figure C-8: Available Servers
12. Press Set and then Test Connection to confirm that the connection has been established.
13. Now that the connection Homer to the LTI database on swordfish has been established, we must activate the connection before launching HLATools. After selecting Homer from the Connections list, choose the Make Active
option from the right-click menu. This activates the Homer connection as shown in Figure C-9.
Figure C-9: Activating a Server Connection
14. When Quimby next launches LTI from his computer (skipjack), he will be able to access the LTI database on the
target server (swordfish) via the Homer connection. You can access remote databases on any computer on which
you have been added to the authorized users list, and you can also add and delete users from the remote computer’s
authorized users list. This administrative function is described below in Editing the Authorized Users List from a
Remote Server, p. 146.
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15. Quimby can re-access his local LTI database, by launching the Connections tool and selecting File > Connection >
Quick Connections > Local to restore active status to the Local connection or by using the Ctrl + L key-stroke
combination. Note that a Ctrl + <…> key-stroke combination is assigned to each Quick Connection. This allows
you to switch between database connections without using the menu. In addition to the default Local Quick Connection (Ctrl + L), you can create up to nine additional Quick Connections (Ctrl + F1 through Ctrl + F9).
Figure C-10: Reactivating the Local Serves Connection
16. To summarize, the controls, the File menu options and the right-click menu options in the Connections dialog have
the following functions:
Add – creates a new connection (Figure C-6).
Apply – saves the settings without closing the dialog box; this allows you to test the connection without leaving the
dialog box; enabled only when a connection has been selected.
Custom Connection – creates a connection for one-time use without saving the connection.
Delete – right-click menu option deletes the selected connection.
Make Active – right-click menu option activates the selected connection.
Modify – accesses the Database Settings dialog (Figure C-7) where you can specify the type of authentication
employed by the connection (SQL or Windows) and define a new connection.
Quick Connections – accesses the local database and up to nine other predefined connections.
OK – saves the settings and closes the dialog box.
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Test Connection – provides confirmation that database settings are valid.
Editing the Authorized Users List from a Remote Server
Once you have established a connection between your computer and a second server, you can also use the User
Management tool to add and remove users from the second server’s list of users authorized to access its LTI database.
Assuming we are using Quimby’s computer of the previous example (skipjack), which we will now call the Remote
server, we can edit the user’s list on the target server (swordfish) as follows:
1. Launch the User Management tool. By default, the users authorized to access the local LTI database are listed in
the main panel. The names of the local server and the SQL server instance are displayed at the top of the panel.
Figure C-11: Listing Local Server Users
2. To change the connection to the target server, select File > Connection > Quick Connections > Homer.
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Figure C-12: Connecting to a Remote Server
3. Once the Homer connection has been made, the users for Homer’s computer appear in the display field just as
though you were observing the list while logged into that computer (see Figure C-5).
4. With access to the target server (swordfish) established, it is possible to add and delete users on the target server
remotely in the same way as one modifies the user list when logged in locally.
Linking to another Server using SQL Authentication
As an alternative to using Windows network security, it is also possible to use SQL authentication to control access
between servers. In SQL authentication access is granted by using a username/password combination that is different
from the Windows username/password.
Important Note: By default, MSDE SQL Server 2000 installation sets the login authentication to Windows authentication mode. The authentication mode can be changed after installation to SQL mode or mixed mode (both SQL and
Windows) by using the SQL Server Enterprise Manager or by appropriately editing the registry on all servers which are
to operate in a peer-to-peer configuration. Changes in authentication mode should be undertaken only by a systems
administrator.
1. Add an SQL user to the another computer’s user list. (This can be done using the User Management tool locally or
remotely.) Select Add SQL User and enter a username/password combination in the Create New SQL User
dialog (Figure C-12).
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Figure C-13: Creating a New SQL User
2. Launch the Connections tool and create a new connection to the other server using the SQL username/password
combination. In this case, we will call the new connection Bart and link to the server named starfish.
Figure C-14: Creating a New Connection for SQL Authentication
3. Enter the same username/password combination used when creating the SQL user on the starfish server
(Figure C-14).
4. Click Test Connection to verify the connection and Set to commit the settings and exit the dialog. This SQL user
will maintain access to the LTI database on the specified server until the connection is deleted from the Connection Names list in the Connections tool dialog.
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Setting up HLATools in a Client/Server Network
To install HLATools in a client/server configuration where Windows Enterprise Network with Domain Based Security
is being used should follow these steps:
1. Log on with Administrative access.
2. Install HLATools Software.
3. Reboot the computer once.
4. Log on again with Administrative access.
5. Provide SQL access privileges to user.
6. Log off as administrator.
7. Log on as User.
8. Continue with the actual installation of the software.
HLATools Security Modes
Three security modes have been implemented to control access to HLATools. In all cases a record of login attempts can
be kept, with username, time and date of login being saved to a log file accessible only by users with administrative
privileges. In all three modes, access to the local computer on which the HLATools suite is installed requires a normal
Windows login to boot up the computer.
•
None – any user can access HLATools without further login.
•
Windows – a user must log in to HLATools using his or her standard Windows username/password combination.
This prevents one user from running HLATools under another user’s account.
•
Local – any user can access HLATools by entering the same string for both username and password in the
HLATools login dialog. The laboratory management can inform users as to the username/password format. The apparent need for a conventional username/password combination should dissuade passersby and the idly curious
from accessing the application. Users should be aware that usernames are included in analysis reports.
The security mode employed on a given computer can be set using the HLATools Configuration utility.
To set or change a security mode setting:
Select Start > Programs > One Lambda > HLATools Configuration Suite > Configuration to access the HLATools
authentication options (Figure C-15).
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Figure C-15: HLATools Configuration
Any change made to the settings will be in effect the next time you launch LTI.
Sharing Luminex Files on a Network
In a typical laboratory workflow, Luminex 100 output files are stored on a computer attached directly to a Luminex 100
Analyzer. The output files are often transferred to a second computer for analysis using HLA Tools (Figure C-16).
Alternatively, the HLA Tools user can read the Luminex output files directly from the Luminex 100 PC. In this section
we shall discuss how to set the file sharing properties to allow direct access to the Luminex output files.
The output files are typically stored on the Luminex PC in a folder called C:\MyBatches\Output. User groups or
individual users can be granted Read permissions for this folder to enable them to read or copy the output files.
In the Windows environment, the root directory is accessible only to administrators. The folder C:\MyBatches\Output
is one level above the root directory, so it can be made accessible to non-administrators if we set the permissions
properly.
Windows 2000 and Windows XP allow you to set user permissions on a per-folder or a per-file basis. Setting the
permissions for the folder automatically sets the same permissions for all files and subfolders. It is also possible to set
more restrictive permissions for a contained subfolder or file. (The following discussion reflects the Windows XP
environment. Terminology for Windows 2000 may differ slightly.)
To set the permissions on the C:\MyBatches\Output folder on the Luminex 100 PC:
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1. Navigate to C:\MyBatches.
2. Right-click on the Output folder and select Properties > Sharing (Figure ).
3. The default setting for most folders is Do not share this folder. Select the Share this folder option.
Figure C-16: Luminex Output Files Workflow
4. The default user group specification in the Share Permissions interface is generally Everyone. Permissions levels
are:
•
Full Control – allows the user to change file permissions, take ownership of files, and perform the actions
granted by the Change and Read permissions.
•
Change – allows the user to overwrite files, change file attributes, and view file ownership and permissions
•
Read – allows the user to read files in the folder, view file attributes, ownership and permissions
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Figure C-17: Output Folder Properties – Sharing
5. Select the Everyone group icon and clear the Full Control and Change checkboxes, making sure the Read checkbox is checked. Now everyone will be able to read or copy Luminex output files from this folder, but not change or
delete them.
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Figure C-18: Share Permissions for C:\LuminexFiles
To simplify access to the C:\MyBatches\Output folder, map this folder as a drive from the HLA Tools PC.
1. Before you start, determine the network identification of the Luminex 100 PC:
•
On the Luminex 100 PC, select My Computer > Properties > Computer Name. (This latter tab is called
“Network Identification” on Windows 2000.)
•
Note the Full Computer Name and the Domain.
2. On the HLA Tools PC:
•
Select My Computer > Map Network Drive …
•
From the Drive: pull-down menu, select an unused drive letter.
•
Select Browse > My Networks Places > Entire Network > Microsoft Windows Network > <network
branch>. (If you do not know the name of the network branch where the Luminex 100 PC is located, ask your
system administrator.)
•
Open the node with name of the Luminex 100 PC, select the Output folder, and click OK to map the drive.
The Output folder will now appear in the listing of Network Drives when you open My Computer on the
HLA Tools PC. (Figure C-19)
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Figure C-19: Mapping C:\MyBatches\Output on a Remote Computer
You will now be able to browse directly to the Luminex 100 PC’s Output folder from within the Lambda
Explorer (Figure 4-1 and Figure 4-2).
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Glossary
This glossary contains terms that may be helpful for beginning users of HLATools™ LABType Interactive. A general
glossary of terms in HLA, immunology, and related fields compiled by One Lambda can be found at
http://www.onelambda.com/resources/glossary.asp.
Glossary Entries
A260/A280 Ratio
A measure of the purity of DNA in a sample can be obtained by comparing the ratio of
spectrophotometric absorbance of the sample at 260 nm to that of 280 nm. Nucleic acid
samples have a higher absorbance at 260 nm than at 280 nm, while protein samples have a
lower absorbance at 260 nm than at 280 nm. It has been empirically determined that a pure
nucleic acid sample has an A260/A280 ratio of 2.0, while a pure protein sample has a ratio
of 0.57. Using this ratio as a guide, one can estimate the relative concentration of nucleic
acid to protein is in a sample. The One Lambda Micro SSP protocol calls for a sample
A260/A280 ratio of between 1.65 and 1.80 which corresponds to a DNA percentage
between 65% to 80%.
ACD
Acid citrate dextrose – a blood anticoagulant used in preparing samples for PCR.
Allele Name
Extensions
In 2002 the WHO Nomenclature Committee for Factors of the HLA System decided that
in cases where the total number of allelic subtypes exceeds 99, a second number series
would be used to the extend the first one. This nomenclature has been now been implemented for several alleles: the extension of the B*15 type (or family) is B*95, and the
extension of the A*02 type is A*92. These nomenclature extensions are observed by LTI.
See http://www.ebi.ac.uk/imgt/hla/nomen_changes.html.
Amino Acid Codes
The following Amino Acid Codes are recognized by the International Union of Biochemistry (IUB) and the International Union of Physical and Applied Chemistry (IUPAC).
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Table Gl-1: IUB/IUPAC Amino Acid Codes
Code
Three-Letter Abbreviation
Description
A
Ala
Alanine
B
Asp, Asn
Aspartic Acid, Asparagine
C
Cys
Cysteine
D
Asp
Aspartic Acid
E
Glu
Glutamic Acid
F
Phe
Phenylalanine
G
Gly
Glycine
H
His
Histidine
I
Ile
Isoleucine
K
Lys
Lysine
L
Leu
Leucine
M
Met
Methionine
N
Asn
Asparagine
P
Pro
Proline
Q
Gln
Glutamine
R
Arg
Arginine
S
Ser
Serine
T
Thr
Threonine
V
Val
Valine
W
Typ
Tryptophan
X
Xxx
Unknown
Y
Tyr
Tyrosine
Z
Glu, Gln
Glutamic Acid, Glutamine
*
TERM
Termination (Stop) Codon
ASO
Allele specific oligonucleotide typing – also called sequence specific oligonucleotide
(SSO) typing.
Class I MHC
Class I MHC molecules usually display “endogenous proteins”, or peptides derived from
the cytoplasm of the cell. These antigens are typically fragments of viral proteins or tumor
proteins from within the cell itself. Class I MHC molecules are made up of two chains, a
heavy chain transmembrane polypeptide and a light chain beta-2 microglobulin that does
not penetrate the cell membrane. Class I OLI products employ two positive control
probes: bead 13 for exon 2 and bead 32 for exon 3.
Class II MHC
Class II MHC molecules usually display peptides derived from extracellular proteins that
are internalized into phagocytic or endocytic vesicles. These extracellular proteins are
typically fragments of bacteria or viruses that have been engulfed and “processed”, or
broken down, by a so-called “professional” antigen presenting cell such as a macrophage
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or dendritic cell. Class II OLI products employ a single positive control probe: bead 34 for
exon 2.
Figure Gl-1: Class I and Class II MHC Molecules
Antigen
α2
α3
Antigen
α1
β2Μ
α
Class I MHC Molecule
Antigen Presentation to
CD8+ T cells
Cell death detection
α1
β1
α2
β2
α
β
Class II MHC Molecule
Antigen Presentation to
CD4+ T cells
In serological screening, cell death is detected by the entry of a stain into dead cells. A
commonly used staining medium is a mixture of acridine orange (AO) and ethidium
bromide (EB) in a solution of bovine hemaglobin. Acridine orange (a green dye) stains
live cells. Ethidium bromide (a red dye) binds to the DNA of lysed, or dead, cells and
excludes the acridine orange. A very strong positive reaction is indicated when over 80%
of the cells are killed. Reactions are graded using the NIH scoring scheme:
Table Gl-2: NIH Reaction Scores
NIH Score
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Cell Death Rate (Reaction Strength)
0
None
1
1 < 20%
2
20 < 40%
4
40 < 60%
6
60 < 80%
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NIH Score
Cell Death Rate (Reaction Strength)
8
80 < 100%
x
unreadable
Some typing sera register weak or moderate reactions if they react with an antigen in the
same cross reactive group (CREG) as the main specificity.
The 0 - 8 scoring convention is also used in some non-serological testing applications. For
example, in LABType Interactive a positive test bead reaction is indicated by an 8, a
negative reaction by a 1.
Clean Database
The Clean Database utility creates the original instance of the LTI database. It must be run
before LTI can be expected to work. Until it is run, there is no database for LTI to use. A
database consists of two parts, the Schema, and the Data. Clean Database builds the SQL
Database Schema, then fills it with some initial values (list of One Lambda products,
serology table values, etc.). Once this has been done, there is enough of a framework so
that the user can import .csv files which contain data to be analyzed, store the results of
analysis, and ultimately create analysis reports.
Close Reaction
When the combined reaction patterns of a pair of alleles are only one false positive or one
false negative removed from yielding a Matched Reaction Pair (MRP), this is termed a
Close Reaction. An example is shown in Figure Gl-2. Here the combined reaction patterns
of alleles DPB1*1401 and DPB1*1301 would be consistent with the positives in the
sample reaction pattern (Rxn) if the sample had also exhibited a positive reaction of
bead 12.
Figure Gl-2: Close Reaction
Code definition
The NMDP code definition is the expansion of the code to show all the component alleles.
For example, the definition of A*33DWH is A*3301/A*3303/*3304/*3305/*3306.
Complement
A group of proteins in the serum that is activated by the antibody-antigen complex on a
cell surface. The result may be the destruction of the antigen or lysis of the presenting cell.
Count Type
The Luminex output files contains two different count data points.
CREG
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•
Trimmed Count – the count for each bead is reduced by a specified percentage; the
default value is 10%.
•
Count – the full count of each type of bead drawn from a well; this is the default
option used during the LTI analysis.
Cross REactive Group – antigens with shared Epitopes.
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EDTA
Ethylene Diamine Tetraacetic Acid – a chelating agent used as a buffer in preparing blood
samples.
ELISA
Enzyme-Linked ImmunoSorbent Assay – the antibody detection method used by the
Lambda Antigen Tray (LAT) assay. The LAT assay is based on the principle of presenting
purified HLA antigens as targets for the binding of a patient’s antibody. In LAT, defined
amounts of purified HLA antigens are presented in the different wells of a Terisaki tray.
The specific binding of serum antibody with any of these antigens is detected by the
subsequent incubation with alkaline phosphatase-conjugated antibody that recognizes
only human IgG. A quantitative measure of the extent of reaction is obtained by spectrophotometric determination following the addition of an enzyme Substrate appropriate for
the development of color. Qualitative assessment of antibody specificity is performed by
the analysis of the LAT reactivity pattern.
Epitope
A unique shape or marker on an antigen’s surface that triggers a corresponding antibody
response. Epitopes are classified as private or public.
•
Private epitope – an individual HLA specificity
•
Public epitope – shapes or markers that many antigens have in common as a result of
having shared amino acid sequences. A single antibody may react to many different
HLA antigens if they share the same amino acid sequences. Antigens with shared
epitopes are called cross reactive groups (CREGs).
False Negative
A bead reaction “missing” from the sample observations. An actual false negative is most
often due to the sample reading being under the cutoff value. When one compares the
negative reading to other sample data and to QC data, one may conclude that it is justified
to adjust the cutoff value downward to allow the negative reading to be reclassified as
positive.
False Positive
A positive bead reaction that is not expected to be positive. False positives are often the
result of well contamination or an error during the preparation of the sample, particularly
during the PCR phase, with the result that the sample reading is over the cutoff value.
When one compares the positive reading to other sample data and to QC data, one may
conclude that it is justified to adjust the cutoff value upward to allow the positive reading
to be reclassified as negative.
False Reaction
After LTI searches for allele pairs with combined reaction patterns that yield exact
Matched Reaction Pairs (MRPs), it searches for allele pairs whose combined reaction
patterns differ by one bead reaction (either positive or negative) from yielding an MRP.
Such an allele pair may be referred to as a “one-away”.LTI records the allele pair and the
bead ID of the bead involved in the false reaction. LTI then searches for allele pairs that
are “two-aways”, “three-aways”, and so forth. This search mode can be invoked by
changing bead cutoff values globally in the Bead Analysis View or locally in a sample’s
Reaction Pattern Histogram.
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Genetic Code
The amino acids are encoded by the following nucleotide triplets. See Table Gl-1for the
full names of the amino acids.
Table Gl-3: Amino Acid Nucleotide Triplets
T
C
Phe (F)
T
A
G
Tyr (Y)
Cys (C)
Ser (S)
Leu (L)
Term (*)
Term (*)
A
Trp (W)
G
T
His (H)
C
Leu (L)
Pro (P)
Arg (R)
Gln (Q)
A
Ile (I)
Asn (N)
Ser (S)
Lys (K)
Arg (R)
Val (V)
Ala (A)
Gly (G)
Glu (E)
A
T
C
A
G
T
Asp (D)
G
C
G
Thr (T)
Met (M)
T
C
C
A
G
Immunomagnetic
beads
Immunomagnetic beads are superparamagnetic particles with monoclonal antibodies
coupled to their surface. The beads can be collected using a magnetic field. When the field
is removed, the beads do not retain any residual magnetism (which is one property of
paramagnetism; the term “superparamagnetism” indicates that the beads display paramagnetic behavior at a lower temperature than would normally be expected;
superparamagnetism tends to become more pronounced as the size of an object diminishes). They can be repeatedly magnetized and redispersed. The specificity of the coupled
monoclonal antibody determines the type of cell collected. The One Lambda FluoroBeads
products employ immunomagnetic beads.
JPN Rank
Demographic information on allelic representations in ethnic Japanese. In LTI, the Japan
Rank A is the most restrictive while Rank ABC is the least.
Local Code
When no NMDP ambiguity code is available to designate a group of allelic subtypes with
a common reaction pattern, LTI designates the group with a temporary local code, usually
of the form XXn. As soon as an appropriate NMDP code has been defined and the internal
LTI table of NMDP codes as been updated, the local code is replaced by the NMDP code.
Log Files [.ldf]
Log files hold all of the log information used to recover the database. There must be at
least one log file for each database, although there can be more than one. The recommended file name extension for log files is .ldf. See also Primary Data Files [.mdf].
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Matched Reaction
Pair
A pair of alleles whose joined bead reaction patterns match that of the observed sample
reaction pattern. If the patient is heterozygous for the sequence being typed, each of the
two alleles will come from a different parent. If the patient is homozygous, the alleles
from each parent will be identical. In various LTI tables, homozygous MRPs are indicated
by an entry only in the Allele 1 cell.
Mean and Median
The Luminex output files provides a number of different quantities that can be used as
“average” readings for the beads.
•
Mean – the mean is the average calculated using all the data points for each bead. For
the dataset consisting of the five data points, 3, 5, 6, 7, 14, the mean would be 35/5
or 7.
•
Median – the median is the value of the data point which separates the upper half of
the data set from the lower half. If there are an even number of data points in the set,
the median is typically calculated as the mean of the two middle values. For the data
set cited above, the median would be 6.
•
Trimmed Mean – the mean is calculated after a specified percentage of the high and
low readings for each bead are thrown out to reduce the influence of outliers; the default value for trimming in a Luminex output file is 5% each from top and bottom. For
the data set cited above, trimming the top and bottom values would yield a trimmed
mean of 18/3 or 6.
•
Trimmed Median – the median bead reading determined after trimming a given percentage off the ends of the data set. For the data set cited above, the trimmed median
will be 6. The trimmed median and the median are generally identical.
Merge Database
The Merge Database function combines two LTI databases and, in the process, archives
both individual databases. If both databases contain the same batch, the two instances are
kept separate since each batch in the merged result is assigned a different internal identification number. If you merge databases on a regular basis, Lambda recommends that you
perform the merge whenever the “from” databases contains a thousand or more samples.
MIC
MHC Class I Chain-related markers appear to play a role in graft rejection. MIC proteins
are single chain polymorphic proteins. Currently (6/2006) there are 51 variants and
approximately 60 recognized alleles. Since MIC gene products are expressed on
endothelial cells, but not on lymphocytes, conventional cross-match procedures do not
detect anti-MIC antibody.
N80/K80
HLA-Cw4 and -Cw6 belong to one allotype group of HLA-C proteins characterized by the
presence of K80 (lysine at the 80th residue) whereas -Cw3 and -Cw7 belong to the other
group that has N80 (asparagine at the 80th residue).
Negative Control
A control intended to provide a reference value for no reaction whatsoever. Negative
control values are typically used to confirm that the data collection methodology was
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sound, to correct for background noise or contamination, and to normalize raw data
readings. The actual nature of a negative control may vary from product to product.
•
In LABType – a probe that will not bind to any HLA sequence.
•
In LABScreen – a negative control serum that contains no HLA antibody according to
LABScreen testing and negative control beads with no antigen attached
•
In Micro SSP – a negative control reaction tube that detects the human beta-globin internal control PCR amplification product; this provides a background value since the
beta-globin gene amplification product is present in every well and is the most likely
PCR contaminant.
NIH Score
A scoring scheme in which serological reactions are assigned values ranging from 0 (no
reaction) to 8 (very strong positive). See Table Gl-2 for a tabulation of scores and their
significance.
NMDP code
An ambiguity code currently consisting of from two to four alphabetic characters. The
characters in an NMDP code correspond uniquely to a concatenation of numbers used to
designate allelic subtypes that share a common reaction pattern. For example, the NMDP
code CKB corresponds to the numbers 01/03/07. Thus the ambiguity code B*45CKB is a
shorthand designation for the allelic subtypes B*4501, B*4503 and B*4507.
The assignment of NMDP codes is quasi-random, with new codes being created as longer
or different numerical concatenations are required. For example:
KBY -> 01/02/03/04/05/06/07/08/09/10/11/12
HDY -> 01/02/03/04/05/06/07/08/09/10/11/12/13
FYH -> 01/02/03/04/05/06/07/08/09/10/11/12/13/14
An example of four-digit allelic subtypes including one null allele would be
ASKK -> 0301/0303N/0304/0306/0307/0308/0309
A full listing of current NMDP codes can be found in the NMDP > View which is
accessed via the HLATools Configuration Suite. For an explanation of allele nomenclature, see the FAQ topic, Alleles – How are they designated?
Normalization
Test reactions are expressed as a percent of the normalized raw reading according to the
formula:
( Raw – NC )
Data = 100 ------------------------------( PC – NC )
(Gl-1)
where
Data = normalized fluorescence reading (shown in most tables)
Raw = raw fluorescence reading value
PC = positive control value
NC = negative control value
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Nucleotide Codes
The following nucleotide codes are recognized by the International Union of Biochemistry
(IUB) and the International Union of Physical and Applied Chemistry (IUPAC).
Table Gl-4: IUB/IUPAC Nucleotide Codes
IUB / IUPAC
Meaning
Description
A
A
Adenine
B
C or G or T
Not Adenine
C
C
Cytosine
D
A or G or T
Not Cytosine
G
G
Guanine
H
A or C or T
Not Guanine
K
G or T
Keto
M
A or C
Amino
N/X
A or C or G or T
Unknown
R
A or G
Purine
S
C or G
Strong
T/U
T/U
Thymine/Uracil
V
A or C or G
Not Thymine/Uracil
W
A or T
Weak
Y
C or T
Pyrimidine
Null Allele
Comparative studies between serology and molecular typing for HLA-A and B loci have
discovered alleles detected by DNA typing but not detected serologically. These HLA
expression variants, commonly referred to a “null” (N) or “low expression” (L) alleles, are
not expressed on the cell surface or are expressed to a very low degree. They are designated by an N or L suffix.
PBS
Phosphate Buffered Saline – a non-toxic saline solution used as a diluent in sample
preparation.
PCR-SSP
PCR Sequence Specific Primer – in PCR-SSP oligonucleotide primers are designed to
obtain amplification of specific alleles or groups of alleles. The typing method is based on
the premise that a completely matched primer will be more efficiently used in the PCR
amplification than a primer with one or more mismatches. The One Lambda Micro SSP™
product employs PCR-SSP analysis.
PCR-SSP has a number of advantages over other PCR-based typing methods:
1. High degree of resolution, with each primer pair defining two linked, cis-located polymorphic sites, which facilitates the typing of heterozygous individual.
2. Results are easy to interpret.
3. The inefficiency of the Taq polymerase in extending mismatched primers is a more
precise reaction than hybridization with ASOs.
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4. The post-amplification analysis is more rapid and simpler than other PCR-based
methods because the typing specificity is part of the PCR.
5. Inexpensive and versatile (it can be used for all Class II loci).
6. For higher resolution, a second analysis with additional pairs of primers can be performed.
Positive Control
Control beads intended to provide a reference value for a complete reaction. Positive
control values are typically used to confirm that the data collection methodology was
sound and to normalize raw data readings. The actual nature of a positive control may vary
from product to product.
•
In LABType – a probe that will bind to all sequences for the particular HLA locus being recognized in the hybridization reaction.
•
In LABScreen – none; positive reactions are defined in terms of cutoff values that are
determined in different ways for the PRA, Mixed and Single Antigen products.
•
In Micro SSP – a primer pair that amplifies a conserved region of the human betaglobin gene; the amplification product of the control primer pair verifies the validity
of the PCR amplification.
Positive Reaction
A normalized value for a bead reaction that meets a preestablished cut-off value.
Primary Data Files
[.mdf]
The primary data file is the starting point of the database and points to the other files in the
database. Every database has one primary data file. The recommended file name extension
for primary data files is .mdf. See also Log Files [.ldf]
Recognition Site
A DNA sequence that is matched by the probe on a bead. The recognition site will usually
be the exact reverse complement of the oligonucleotide sequence of the probe. Recognition sites are described in terms of the amino acids which the nucleotides encode rather
than in terms of the nucleotides themselves.
Consider the recognition site entry for bead #1 in Figure Gl-3:
5----Y-----14
MRYFFTSVSR
(probe)
(reference sequence)
This entry indicates that the probe is specific for the nucleic acid sequence that encodes
the amino acids in the pertinent locus at positions 5 through 14 with a substitution of
tyrosine (Y) for phenylalanine (F) at the 9th position. (See Table Gl-1above for a listing of
amino acid codes.) The reference sequence alignments for the various loci can be found at
the Anthony Nolan Research Institute website under the category of HLA Class I and II
Sequence Alignments. See http://www.anthonynolan.org.uk/HIG/data.html.
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Figure Gl-3: Raw Data Table – Recognition Sites
RFLP
Restriction Fragment Length Polymorphism – the first molecular typing method applied
to the Class II MHC (in 1982). RFLP analysis is based on restriction endonuclease
cleavage at polymorphic restriction sites. These sites, specific for each enzyme (endonuclease) used, are located in both coding and non-coding regions of the genes. The strong
linkage disequilibrium between specific polymorphic restriction sites and coding sequence
variations allows RFLP analysis to be useful in HLA Class II DNA typing. A good correlation exists between DNA-RFLP analysis and phenotypic typing for Class II specificities.
Early forms of RFLP analysis using radio-tagged probes have been largely replaced by
PCR-based analytical methods.
Secondary antibody
In ELISA assays, the “detection” antibody that binds to the serum antigen which has
already bound to the purified “capture” antibody attached to the bottom of the well. The
secondary antibody in turn becomes bound to the colorimetric substrate. The strength of
the signal from the substrate can be used to make a quantitative determination of the
amount of antibody in the sample.
SSO
Sequence Specific Oligonucleotide – SSO typing involves PCR amplification of a chosen
sequence using primers flanking the sequence. The amplified DNA is immobilized on a
membrane and hybridized with selected oligonucleotide probes that are labelled with
radioactive 35S or 32P, by ELISA, or by chemical markers.
Substrate
In ELISA assays, the substrate is the chromogenic chemical species that produces a signal
when it binds to the secondary antibody. One Lambda ELISA assays typically use either
pNPP (para-nitrophenyl phosphate) or BCIP (5-bromo-4-chloro-3-indolyl phosphate) as a
substrate.
Unicode
A standard for representing characters as integers. Unlike ASCII, which uses 7 bits for
each character, Unicode uses 16 bits, which means that it can represent more than 65,000
unique characters.
Update Database
This utility rebuilds the LTI database with new product information when the user updates
the HLATools software. In contrast to Clean Database which creates an entirely new
database, Update Database maintains all the analysis results and user input that was
contained in the old database.
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IIndex
Symbols
/ – slash
alternative allelic subtypes, 73
~ – tilde
range of allelic subtypes, 73
self-evident filepath name, 91
A
A*02/A*92 allele family, 155
A1 4Digit\A2 4Digit, 38
A260/A280 Ratio, 155
ABDR locus reports, 6
Accept function
re-enabling, 47
requiring review by two analysts, 47, 108
ACD – acid citrate dextrose, 155
Active content
disabling browser warning message, 10
Allele name extensions, 155
Ambiguity codes
local, 120, 160
NMDP, 120
Amino acid codes, 155
AO – acridine orange, 157
Application Data
hidden folder, 20
ASO – allele specific oligonucleotide, 156
Authentication mode
SQL, 139, 147
Windows, 139
Authorized users list
editing, 146
B
B*15/B*95 allele family, 155
Batch analysis
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BCIP, 165
Bead Analysis Histograms
viewing entire width, 109
Bead Probe Information
data sheet, 73
Bead specificities
copying to clipboard, 54
displaying, 54
Bw4/Bw6 serotypes, 77
C
Cell death detection, 157
Character encoding type
specifying, 109
Class I/II proteins, 156
Clean Database
need to run, 27
using all.cmd script, 136
Close reactions
illustrated, 158
Code point, 109
Codes
amino acids, 155
Local – XXn format, 46, 120
NMDP, 120, 162
nucleotide, 163
nucleotide triplets, 160
Color coding
in Raw Data table, 63
in Reaction Grid, 57
in Sample Data bead profiles, 40
Comments Editor
HLATools – Comments Editor tool, 47, 107
Complement, 158
Corrected typing
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same as Accepted typing, 51
Count Type, 71, 158
CREG – cross reactive group, 158
Cutoff Sensitivity, 40, 71
D
Data sheets
Bead Probe Information, 73
Resolution Limitations, 73
Data transfer file export type, 108
Data Type
means and median, 71
Database
Log Files, 160
Primary Data Files, 164
Database Login
when printing reports, 116
Date formats
changing, 107
E
EB – ethidium bromide, 157
EDTA – ethylene diamine tetraacetic acid, 159
ELISA, 165
Epitopes – private and public, 159
Excluded beads
background color in Reaction Grid, 40, 57, 63
Export type
Comments editor, 108
data transfer file, 7, 108
F
False Negative
defined, 159
False Positive
defined, 159
False reactions, 159
Figures
see also Tables
All Alleles Table, 51
generating, 124
Allele Symbols in the Reaction Pattern Histogram, 50
Assignments View w/ Close Reactions, 49
Assignments View w/ MRPs, 45
Attach/Detach Database Tool, 130, 131
Backup Database, 23
Batch Results Table, 36
Bead Analysis View, 39
Bead information – where to find, 72
Character Encoding Type – specifying, 150
Class I and Class II MHC Molecules, 157
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Clean DataBase Log Message, 17
Close reaction – example, 158
Close reactions sorting options, 106
Closest Reactions Tables, 53
Comments Editor utility, 108
Connections Tool, 142
Control Values – Minimum Bead Count, 43
Control Values – Positive Control Values, 42
Custom Reports
Advanced Search Filters, 85
Column Search Filters, 84
Patient Reports search options, 82
Patient Search Options, 83
Samples to Report On, 84
Customer Information Dialog, 15
Database – authorized users, 141
Database settings, 143
Database settings – creating, 143
Export Type – specifying, 150
Flowchart – Updating HLATools, 21
HLATools data reference files, 70
Installshield Modify … Message, 14
LABType Configuration Panel, 109
LABType Product Information, 71
Lambda Explorer
Adding a Folder, 28
File Tree, 28
Local server – reactivating connection to, 145
Log-on type – specifying, 150
LTI Batch/Sample File Tree, 34
Luminex Files folder – specifying, 150
Microsoft .NET Framework Installation, 14
MRP tabulation, 123
MRPs from “Golden” alleles, 121
NMDP code definitions – where to find, 76
Patient Information Subview, 55
Patient Typings Table, 68
Probe recognition sites, 165
Product Information Controls, 70
Product Update
downloading LABType product information, 74
missing file extension, 75
Product/Lot
Selection, 30
Program Maintenance, 18
Raw Data
double column sort, 65
low bead count, 64
low readings, 64
sort by minimum bead count, 64
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sort by recognition site, 63
Reaction Grid
“Golden” allele, 121
bidirectional sort by allele pattern, 62
clustering positive reactions, 60
default view, 120
False Negative, 59
False Positive, 59
restoring allele sort order, 61
restoring bead sort order, 60
sort fields, 61
sorting by bead usage, 59
Rearranging table columns, 106
Report header customizing
company icon, 81
laboratory information, 81
Report Header Laboratory Information, 80
Report Templates folder – specifying, 150
Runtime Error Message, 19
Sample Reaction Pattern Grid, 56
Sample/Product Mismatch, 30
Selecting a File for Import, 29
Serological equivalents – where to find, 77
Server – activating connection to, 144
Server – listing available …s, 144
Server – listing local users, 146
Server – Quick Connection to remote …, 147
Server connection – activating, 144
Setting Database Name in ALL.bat, 131
Setup Options – Custom, 19
Simple Network Installation, 139
Specificity Subview, 54
SQL authentication, 148
SQL Server and Database Status, 22
SQL Server Network Utility, 136
SQL user – creating new, 148
Synopsis of Batch Analysis, 33
Synopsis of Sample Analysis, 44
Type/Subtype Tables, 52, 126
Updating LABType product information, 73
User Management Tool, 139
Windows network domain users, 140
Windows XP
changing security policy, 114
modifying firewall settings, 113
Firewall
making a port through, 112
G
Genetic code, 160
“Golden” allele, 58, 60, 121
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HLA allele nomenclature, 116
HLATools
installing new version …, 20
LABType Home, 5
LABType Interactive (LTI)
described, 5
Lambda Explorer, 5
Lambda Reporter, 6
Patient Explorer, 6
HLATools Configuration Suite
Attach/Detach Database, 7
Backup Database, 7
Clean Database, 7, 158
Comments Editor, 6
complete uninstall of HLATools, 20
Connections Tool, 7
HLA Update, 6
System Diagnostics, 7
Update Database, 165
User Management, 8
I
Immunomagnetic beads, 160
Installation – updated versions of HLATools, 20
Internet Explorer security options, 10
J
JPN rank, 160
L
Lab Information Interface, 81
LABType Classic
importing analysis results into HLATools, 132
Local code, 160
Low expression alleles, 163
LTI database – showing currently attached …, 7
Luminex files
C:\LuminexFiles folder, 16, 150
C:\MyBatches folder, 150
setting sharing permissions, 150
M
Mean and Median – defined, 161
MIC, 161
Microsoft .NET Framework, 13
Microsoft SQL Server service
stopping, 20
MRP – matched reaction pair, 161
MRP detection flowchart, 119
MSDE – Microsoft SQL Server Desktop Engine, 13, 20, 114,
116
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MSDE version and product level, 115
N
N80/K80 – C-locus allotypes, 161
Named Pipes, 116, 136
Negative control, 161
Networking protocol versions, 115
NIH Score, 157, 162
NMDP allele codes
code definition, 158
examples, 162
listing, 6, 75
revision dates, 75
scheduling automatic update, 128
Normalization formula, 162
Null alleles, 163
P
Patient Explorer
purpose of, 67
Patient IDs, 67
trivial, 31
PBS – Phosphate Buffered Saline, 163
PCR-SSP – PCR Sequence Specific Primer, 163
pNPP, 165
Positive controls, 164
Class I and Class II, 47, 156
Positive reaction
defined, 164
Probe ID, 73
Product/Lot
deleting from Product Information table, 71
retired, 73
R
Reaction Pattern Histogram
in analysis report, 98
printing, 41
viewing entire width, 109
Reanalysis of entire batch, 35
Recognition Site, 164
Report formats
A4 page size, 91
template files, 91
US Standard page size, 91
Reports
customizing header information, 80
customizing patient report, 82
overriding default paper size, 90
print and export functions, 92
printing and exporting, 92
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report filters, 93
Resolution Limitations
data sheet, 73
S
Sample IDs
handling duplicate, 31
trivial IDs not matched to patient, 31
Scripts
all.cmd, 136
Security
mode options, 149
modifying policies, 114
Serological equivalents
B–, 77
BBlank, 77
Server Network Settings, 116
running svrnetcn.exe, 135
SQL server – showing current instance, 7
SSO – Sequence Specific Oligonucleotide, 5
SSP – Sequence Specific Primer, 5
Substrate in ELISA tests, 165
Superparamagnetism, 160
T
Tables
see also Figures
Allele sorting in the Reaction Grid, 128
Amino acid codes, 156
Amino acid nucleotide triplets, 160
bead profile color coding, 40
creating the Type/Subtype table, 125
grouping MRP alleles, 122
HLA allele nomenclature, 116
examples, 117
MSDE versions and product levels, 115
NIH scores and cell death rates, 157
Nucleotide codes, 163
raw data table color coding, 63
reaction grid color coding, 57
TCP – Transmission Control Protocol, 112
TCP/IP, 116, 136
Trimmed Count – default setting, 158
Trimmed Mean – default setting, 161
Trimmed Median, 161
U
UDP – User Datagram Protocol, 112, 113
Unicode, 165
Update Database
database does not exist, 112
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execution denied, 112
UTF – Unicode Transformation Format, 109
Utilities
see HLATools Configuration Suite
V
Visible
appearance priority in Product Select, 72, 73
W
Wireless network installation, 135
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