Roche cobas®h 232 Troponin T METHOD AND SAMPLE

Roche cobas®h 232 Troponin T METHOD AND SAMPLE
 Roche cobas® h 232 Troponin T METHOD AND SAMPLE
COLLECTION
1.
PURPOSE AND SCOPE
The purpose of this document is to describe the procedure for performing a
cardiac Troponin T test using the Roche cobas h 232 analyser. The Roche
cobas h 232 analyser can be used by healthcare professionals for measuring
major cardiac blood markers. This document will be focusing on Troponin T,
and further information on the other parameters can be found on their website.
2.
HAZARDS
Patient Samples
All patient samples should be treated as potentially infectious and handled
appropriately. Standard precautions should be employed. Personal protective
equipment should be worn when processing samples, performing
maintenance and troubleshooting procedures.
3.
CLINICAL SIGNIFICANCE
Acute Coronary Syndrome (ACS) is a term used to describe a group of
conditions resulting from insufficient blood flow to the heart muscle. 1 These
conditions range from atypical chest discomfort and non specific
electrocardiographic changes to a large ST-segment elevation, myocardial
infarction and cardiogenic shock.2 Symptoms can include chest pain including
tightness and heaviness in the chest, discomfort in the arms and upper body,
shortness of breath and other constitutional symptoms including sweating,
nausea and light-headedness.1
Diagnosis of ACS is based on a complete medical history, physical
examination, electrocardiogram to evaluate the electrical activity of the heart
and blood tests to evaluate the presence of biological markers resulting from
cardiac cell injury.1
Troponin T & I are members of a group of cardiac regulatory proteins which
function to regulate the calcium mediated interaction of muscle filaments actin
and myosin resulting in contraction and relaxation of striated muscle. 4
Troponin T is almost exclusive to the myocardium, with small amounts
expressed in skeletal muscle not detectable in current Troponin T assays.4
Insufficient blood flow and oxygen supply to the heart muscle causes necrosis
of the myocardium and subsequent release of Troponin T & I into the
bloodstream.5
Troponin T in the bloodstream rises to detectable levels after 4-6 hours, peaks
at 10-12 hours and can be detected for up to 14 days post infarction.5,6
Troponin I is released from necrotic cardiac myocytes into the bloodstream
within hours (~4-8 hours) after the onset of chest pain. The peak TnI
concentration is generally reached in 12-48 hours.7 Troponin I serum levels
Roche cobas h 232 Troponin T Method and Sample Collection IECTnT.1.v1 19.1.12 1
can remain elevated for up to 4–7 days.5
The diagnostic utility of Troponin T & I to detect myocardial necrosis and to
enable risk stratification in patients with ACS is well established.5,8
Furthermore, the use of Troponin T as a prognostic indicator for recurrence of
ischaemic events and death in ACS patients is increasing. 5,9
Results from PoCT devices measuring Troponin T & I should always be used
in conjunction with clinical presentation, history and other diagnostic
information.
4.
TEST PRINCIPLE
The test strip contains two monoclonal antibodies specific to cardiac troponin
T (cTnT) of which one is gold-labelled, the other biotinylated. The antibodies
form a sandwich complex with the cTnT in the blood. Following removal of
erythrocytes from the sample, plasma passes through the detection zone in
which the gold-labelled cTnT sandwich complexes accumulate and the
positive signal is displayed as a reddish line (the signal line). Excess goldlabelled antibodies accumulate along the control line, signalling that the test
was valid. The intensity of the signal line increases in proportion to the
troponin T concentration. The optical system of the cobas h 232 instrument
detects the two lines and measures the intensity of the signal line. The
integrated software converts the signal intensity to a quantitative result and
shows it in the display.
4.1
Interference
No interference was observed up to the following concentrations for all
analytes:
Bilirubin
20 mg/dL
Hemolysis (Hb)
200 mg/dL
Biotin
10 ng/mL
Lipaemia (triglycerides)
440 mmol/L
Rheumatoid factors 300 IU/mL
•
•
•
•
•
•
•
The assay is unaffected by haematocrit values between 30 – 50%
In patients receiving therapy with high biotin doses (i.e. > 5 mg/day), no
sample should be taken until at least 8 hours after the last biotin
administration
High concentrations of lipoic acid (e.g. in pharmaceuticals or as food
additives) can lead to lower measurement values
There is no high-dose hook effect at Troponin T concentrations < 200000
ng/L
Very high concentrations of Troponin T may cause the control line to fail to
appear and the instrument may display an error message.
Patient samples containing heterophilic antibodies may react in
immunoassays to give falsely elevated or decreased results
Strong electromagnetic fields may interfere with the proper operation of
the meter
Roche cobas h 232 Troponin T Method and Sample Collection IECTnT.1.v1 19.1.12 2
Patents:
Important!
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substances
and/or
factors not listed above
US 5,463,467; US It
5,424,035;
US 5,334,508;
5,206,147;
US 5,240,860; US
5,382,523;
US
5,521,060;
US
5,268,269;
US
6,506,575;
US
5,281,395
may interfere with the test and cause false results.
4.2
Accuracy
0123
This product fulfills the requirements for Directive 98/79/EC on in vitro
diagnostic medical devices.
ACCU-CHEK, ACCUTREND, COBAS, SAFE-T-PRO and SOFTCLIX
are trademarks of Roche.
0 5007607001 (01) – 06/07 EN
A comparison of 3 lots of the Roche CARDIAC T Quantitative test with the
Roche Diagnostics
GmbH T test in a clinical patient population showed slopes between
Elecsys
Troponin
D-68298
Mannheim,
0.80 and 1.20 Germany
in the majority of the method comparisons with a correlation
coefficient of ≥ 0.9.
www.roche.com
4.3
Precision
Repeatability was measured with 3 lots of the Roche CARDIAC T Quantitative
tests and heparinised human blood. The majority of the variation coefficients
were below 9 % over the entire measurement range. Intermediate precision
was measured with the Roche CARDIAC Control Troponin T quality control in
5 different hospitals. The majority of the variation coefficients were below 11%.
5.
INSTRUMENT
Product specifications
5.1
Operating Conditions and Technical Data
Temperature range
18o – 32oC
Relative humidity
10 - 85% (non-condensing)
Maximum altitude
4000m
Position
Place meter on a level, vibration-free
surface while applying the sample until the
necessary sample has been absorbed
completely by the test strip
Measuring range
100 – 2,000 ng/L
Sample size
150 µL
Roche cobas h 232 Troponin T Method and Sample Collection IECTnT.1.v1 19.1.12 3
Test time
12 minutes with 2 minutes for sample
detection
Memory
500 test results with date, time and
comments, 500 liquid control results, and
200 code chip records (100 test + 100 QC)
Barcode scanner
Yes
Interface
Infrared interface, LED/IRED Class 1
USB and Ethernet port; printer
Battery operation
Yes – handheld battery pack
(rechargeable)
Mains connection
Yes - Input: 100-240 V (± 10%)/ 50-60Hz
/400 mA, Output: 7.5 V DC / 1.7 A
Number of tests with fully charged
battery
Approx. 10 tests
Safety class
Class III
Automatic power-off
Yes – Programmable 1 - 60 minutes
Dimensions
275 x 102 x 55 mm
Weight
650g incl. handheld battery pack and
scanner
5.2
Storage and transport conditions
Temperature range
Meter (In original container)
-25o to +70oC
Relative humidity
10 - 85% (non-condensing)
6.
SPECIMEN REQUIREMENTS
6.1
Sample Material
Venous whole blood stored in lithium or sodium heparin tubes without
separating gel are acceptable. Blood collection tubes containing EDTA,
citrate, sodium fluoride or other additives are not acceptable.
Samples are stable for 8 hours at room temperature. Do not refrigerate
or freeze samples.
Venepuncture (see suitable anticoagulants above)
• Skin surface must be cleaned with an alcohol swab and dried well prior to
collection to ensure there are no substances on the skin surface
• Ensure sample is properly mixed and at room temperature before testing
• Sample stability: 8 hours at room temperature. Do not refrigerate or freeze
sample.
Roche cobas h 232 Troponin T Method and Sample Collection IECTnT.1.v1 19.1.12 4
7.
CARTRIDGES/REAGENTS
7.1
•
•
•
•
•
Storage and handling
Test strips should be refrigerated at 2° - 8°C. DO NOT FREEZE.
Test strips can be stored up to 1 week at room temperature at 15° - 25°C.
Perform a test at temperatures between 18° – 32°C
Use the test strips at 10 - 85% humidity. Do not store the test strips in high
heat and moisture areas such as the bathroom or kitchen and keep away
from direct sunlight.
Test strips:
o Can be used immediately after removal from the refrigerator
o Must be used within 15 minutes once the pouch has been opened.
o Must be discarded if they are past their use by date. Expired test strips
can produce incorrect results.
o Can be used until the printed use by date when they are stored and
used correctly.
7.2
Storing information about test strips
Every pack of test strips includes a lot-specific code chip which provides
information about the lot-specific properties of the test strip. On opening a new
box of test strips, insert the code chip into the meter. If not inserted when
starting a new lot, the instrument display prompts the user to insert the chip.
To ensure that the code chip and test strip lot match, compare the lot number
in the display with the number on the code chip.
8.
CALIBRATION
The Roche CARDIAC T Quantitative test is calibrated against the
Elecsys Troponin T hs test using serum. The instrument automatically
reads in the lot-specific calibration data from the code chip; thus
operator calibration is not necessary.
9.
QUALITY CONTROL
Quality control material (perform as per your organisation’s
protocol)
Accurately testing known levels of Troponin T ensures that the system
and your technique used in testing give accurate results on patient
tests. The control solutions have defined (known) values. The results for
these solutions must first fall within a certain acceptable range in order
to allow valid patient testing.
A quality control test should be performed every time a new shipment of
test strips are received, when a new lot number of test strips are used, if
the clinical picture does not correlate with the patient test results, after
major maintenance, and at a minimum of once a month.
Roche cobas h 232 Troponin T Method and Sample Collection IECTnT.1.v1 19.1.12 5
Enrolling in an External Quality Assurance Program is encouraged to
objectively compare results with other users using the same method of
testing. If an External Quality Assurance Program is not available,
monthly lab comparisons are encouraged.
The Roche Cobas h 232 uses the following methods for quality:
• Code chips
• IQC (Electronic QC)
• Control solutions
9.1
Coding the meter
• Use the new code chip that comes with every new box of test strips.
• Compare the code number on the chip with the corresponding code
number on the box of test strips.
• Insert the code chip into the code chip slot, located at the top of the meter,
until you feel it snap into place.
NOTE: Do not force the code key into the meter; it only goes in one way.
9.2
Electronic quality control (IQC)
The Roche CARDIAC IQC test serves as a performance check for the
optical system of the cobas h 232 device. The IQC consists of two
Troponin strips with already set positive results (one is a low positive
and one is a high positive). The strips are reusable and test the internal
mechanisms of the instrument to ensure the intensity of the positive line
is read correctly. The IQC should be performed weekly, alternating
between the two levels. Store the IQC strips unopened, at 2-30 °C up to
the stated expiration date. After opening, store for up to 6 months.
•
•
•
•
•
•
Bring the Roche CARDIAC IQC test strip to room temperature before
starting the measurement
From the main menu of the instrument select QC TEST
When prompted remove one test strip from the container and closer the
container immediately.
Insert the IQC strip into the meter (when the instrument asks for a code
chip, insert the code chip from the IQC box).
The instrument will take approximately 20 seconds to perform the test and
when completed the instrument will indicate if the test has passed or
failed.
Remove the test strip (low or high) from the device directly after the
measurement is performed and place it quickly into its container to protect
it from dust and moisture.
Roche cobas h 232 Troponin T Method and Sample Collection IECTnT.1.v1 19.1.12 6
NOTE: Do not touch or wipe the signal line area of the test strip. Do
not apply any sample material to the test strip. Do not expose the
test strip to sunlight.
9.3
Running control solutions
The control solutions have two levels:
• Roche CARDIAC Control Troponin T quality control, level 1
• Roche CARDIAC Control Troponin T quality control, level 2
- each with a lot-specific encoding chip
Store the controls at 2-8 °C and tightly capped when not in use.
The stability of the lyophilized control serum at 2-8 °C is up to the stated
expiration date. Stability of the components in reconstituted control
serum at 2-25 °C is 24 hours and at and below -20 °C is 12 weeks (can
be frozen up to 5 times in the original vial). Frozen or refrigerated
reconstituted control material must be brought to room temperature
prior to use.
Preparing the control solution
• Carefully open a vial, avoiding the loss of lyophilized control serum
• Pipette in exactly 1.0 mL of distilled water.
• Carefully close the vial and dissolve the contents completely by occasional
gentle swirling over 15 minutes.
NOTE: Avoid the formation of foam
Inserting the test strip
• Turn the instrument on by pressing the On/Off button for longer than 5
seconds
• Wait for completion of the self-test
• Touch the QC Test button
• The test strip symbol prompts you to insert the test strip
• Remove the test strip from the foil package
NOTE: Only remove the test strip from the foil package when you are
ready to perform a test.
• Hold the test strip so that the application and test areas are facing up.
Insert the test strip quickly into the test strip guide of the meter using a
smooth, even motion. Slide the test strip in as far as it will go and a beep
tone will indicate that the meter has detected the test strip.
NOTE: If you are using a new test strip lot number and have not inserted
the code chip yet, you will be prompted to do so now.
• If you are using new control material, remove the code chip (for the test
strip), press “New” and insert the code chip that came with the control
material
NOTE: Alternatively, the QC Lot number is stored in the memory, and you
can select the code for your current control material from the list.
• Select the QC level
• The thermometer symbol shows that the test strip is warming up, and the
parameter and code chip number are also displayed on the screen.
Roche cobas h 232 Troponin T Method and Sample Collection IECTnT.1.v1 19.1.12 7
Applying the Control Solution
• When the warming up process is complete, a further beep tone sounds
and a pipette icon appears on the screen
• Using a pipette or syringe apply exactly 150 µL (0.15ml) of control solution
to the application area
NOTE: You have 5 minutes to apply the entire sample to the application
area. Do not use a sample that has air bubbles, and do not touch the
pipette tip to the application zone.
• Touch the tick button to confirm that the sample has been applied. The
meter will now have an hourglass symbol while it detects the sample.
Results
• Once the sample has been detected, the actual measurement starts, and
the countdown will begin.
NOTE: Do not touch the test strip until the result is displayed on the
screen
• The target value and range will be shown on the display along with “Pass”
or “Fail” and automatically stored in the memory.
10.
TEST PROCEDURE
Check the charge level on the screen. If the charge is low, connect the device
to the power supply.
10.1 Code chip
The code chip provides the meter with important manufacturer-specific data
that it needs to perform a Troponin T test. The code chip contains information
about the test method, the lot number and the expiry date of the new test
strips. The meter is ready to use once the code chip has been inserted. See
Roche cobas h 232 Troponin T Method and Sample Collection IECTnT.1.v1 19.1.12 8
section 9.1 for details.
10.2
Performing the Test
Inserting the test strip
• Turn the instrument on by pressing the On/Off button for longer than 5
seconds.
• Wait for completion of the self-test.
• Touch the Patient Test button and enter the Patient ID using the
touchscreen keypad or scan the patient barcode.
•
•
The test strip symbol prompts you to insert the test strip. Remove the test
strip from the foil package only when you are ready to perform a test.
Hold the test strip so that the application and test areas are facing up.
Insert the test strip quickly into the test strip guide of the meter using a
smooth, even motion. Slide the test strip in as far as it will go and a beep
tone will indicate that the meter has detected the test strip.
NOTE: If you are using a new test strip lot number and have not inserted
the code chip yet, you will be prompted to do so now.
Troponin T
Applying the sample
• The thermometer symbol shows that the test strip is warming up, and the
parameter and code chip number are also displayed on the screen.
• When the warming up process is complete, a further beep tone sounds
and a pipette icon appears on the screen
• The meter is ready to perform the test and is waiting for the blood to be
applied.
Roche cobas h 232 Troponin T Method and Sample Collection IECTnT.1.v1 19.1.12 9
•
•
Using a pipette or syringe apply exactly 150 µL (0.15ml) heparinised whole
blood to the application area
NOTE: You have 5 minutes to apply the entire blood sample to the
application area. Do not use a sample that has air bubbles, and do not
touch the pipette tip to the application zone.
Touch the tick button to confirm that the sample has been applied. The
meter will now have an hourglass symbol while it detects the sample.
NOTE: Do not add more blood after the test has begun.
Trop T
TT
Results
• Once the sample has been detected, the actual measurement starts, and
the countdown will begin.
NOTE: Do not touch the test strip until the result is displayed on the
screen
• The result will be shown on the display and automatically stored in the
memory.
• Remove the test strip from the measurement chamber and turn off the
meter by pressing the On/Off button for longer than 2 seconds.
NOTE: A negative result should display 1 line and a positive result should
display 2 lines in the reading window of the test strip. A visual check of the
test strip following the test is recommended to double check the instrument
as if too much blood is applied to the strip it can filter into the reading
window affecting the results. The reading window should remain clear with
only a tinge of pink. If too much blood is added it will turn red and the test
should be repeated.
Trop T
TT
Trop T
TT
120 ng/L
Roche cobas h 232 Troponin T Method and Sample Collection IECTnT.1.v1 19.1.12 10
11.
RESULTS
11.1 Expected values
The measuring range is 100 – 2000 ng/L.
11.2 Interpretation of results
Troponin T
Result
Concentration
Displayed
Below 50 ng/L
Trop T < 50
ng/L
Between 50 ng/L
and 100 ng/L
Trop T
50 – 100 ng/L
Between 100 ng/L
and 2000 ng/L
For example,
Trop T 900
ng/L
Trop T > 2000
ng/L
Above 2000 ng/L
Comment
Acute myocardial infarction not likely,
but still possible; in context of clinical
assessment repeat the
test (e.g. after 3–6 h) to detect rising
Troponin T levels.
Acute myocardial infarction possible,
repeat the test to detect rising Troponin
T levels in context of clinical
assessment according to guidelines;
search for differential diagnosis and
other causes of Troponin T elevation.
Acute myocardial infarction likely;
consider differential diagnosis for other
causes of Troponin T elevation.
Acute myocardial infarction very likely;
consider differential diagnosis for other
causes of Troponin T elevation.
11.3 Transferring Data to a printer or computer
• Using the infrared interface, you can send test results directly to a printer.
• To print the result, align the infrared sensors on both the instrument and
printer and press the printer button.
NOTE: The printer uses thermal paper and will fade over time. Results
should be photocopied and stored in patient’s notes.
• Using the data ports of the handheld base unit (docking station), you can
upload stored test results to a PC/host system (e.g. cobas IT 1000 PoC
data management system).
NOTE: Enabling the connection to a computer disables the connection to
a printer (and vice versa)
12.
MAINTENANCE
•
•
Turn off the meter before cleaning it. Unplug the power supply unit and
remove the handheld battery pack.
First remove any blood and other dirt using water or soapy water then
disinfect the meter
Roche cobas h 232 Troponin T Method and Sample Collection IECTnT.1.v1 19.1.12 11
•
•
Use only the following items for cleaning: ordinary lint-free cotton buds,
lint-free tissues
Suitable cleaning agents include: ammonium chloride solution (2%),
diluted bleach solution (1:10), mild soapy water, Dispatch ®, citric acid
(2.5%), hydrogen peroxide (0.5%), sodium hypochlorite solution (0.6%),
70% isopropyl alcohol, CoaguWipe Bleach Towel (only used for cleaning
the outside of the meter)
Cleaning the Sampling Area
• Remove the sample application cover by pulling it forward horizontally (in
the direction of the arrow).
• In case of significant dirt or contamination, you can rinse the sample
application cover (separately from the meter) under warm running water.
Dry the sample application cover with a fresh tissue.
• Clean the outside of the meter with a lightly moistened tissue. Then dry
the meter with a fresh tissue.
Cleaning Test Strip Guide
• Clean the easily accessible and visible pipetting field area of the test strip
guide with a moistened cotton bud or tissue.
• Dry the test strip guide with a fresh tissue.
NOTE: Do not insert any objects into the concealed areas of the
measurement chamber as this might damage the optical components of
the meter.
• Clean the membrane (small circle) in the visible area at the end of the test
strip guide with a moistened cotton bud or tissue.
• Allow the inside of the test strip guide to dry for about 10 minutes.
• Re-attach the sample application cover to the housing and make sure that
it snaps correctly into place.
Roche cobas h 232 Troponin T Method and Sample Collection IECTnT.1.v1 19.1.12 12
13.
REFERENCES
This method has been adapted from the Roche cobas h 232 System
Operator’s Manual, test strips and control solution package inserts.
1. Torpy, JM Burke, AE & Glass, RM 2010 ‘Acute Coronary Syndromes’, The
Journal of the American Medical Association, vol 303, no. 1, p90.
2. Scirica, BM 2010 ‘Acute Coronary Syndrome: Emerging Tools for Diagnosis
and Risk Assessment’, The Journal of the American College of Cardiology, vol
55, no.14, pp. 1403-15.
3. Chew, DP, Aroney, CN, Aylward, PE, Kelly, A, White, HD, Tideman, PA,
Waddell, J, Azadi, L, Wilson, AJ & Ruta, LM 2011 ‘2011 Addendum to the
National Heart Foundation of Australia/Cardiac Society of Australia and New
Zealand Guidelines for the Management of Acute Coronary Syndromes (ACS)
2006’, Heart Lung and Circulation, vol 20, no. 8, pp. 487-502.
4. Sharma, S, Jackson, PG, Makan, J 2004 ‘Cardiac Troponins’, Journal of
Clinical Pathology, vol 57, no. 10, pp. 1025-6.
5. Daubert, MA, Jeremias, A 2010, ‘The utility of troponin measurement to detect
myocardial infarction: review of the current findings’, Vascular Health and Risk
Management, vol. 6, pp. 691-699.
6. Roche Diagnostics 2011, ‘Roche CARDIAC T Quantitative Troponin T
Quantitative’ Test strip package insert, Mannheim, Germany.
7. Radiometer 2011, ‘Radiometer TnI Test Kit’ Test cartridge package insert,
Bronshoj, Denmark.
8. Keller, T, Zeller, T, Peetz, D, Tzikas, S, Roth, A, Czyz, E, Bickel, C, Baldus, S,
Warnholtz, A, Fröhlich, M, Sinning, CR, Eleftheriadis, MS, Wild, PS, Schnabel,
RB, Lubos, E, Jachmann, N, Genth-Zotz, S, Post, F, Nicaud, V, Tiret, L,
Lackner, KJ, Münzel, TF, Blankenberg, S 2009, ‘Sensitive troponin I assay in
early diagnosis of acute myocardial infarction’, N Engl J Med, vol. 361, no. 9,
pp. 868-877.
9. Waxman, DA, Hecht ,S, Schappert, J, Husk, G 2006, ‘A model for troponin I as
a quantitative predictor of in-hospital mortality’, J Am Coll Cardiol, vol. 48, no.
9, pp. 1755 – 1762.
Roche cobas h 232 Troponin T Method and Sample Collection IECTnT.1.v1 19.1.12 13
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