3-in-1 Flourescence Immunoassay Test kits for Cardiac Troponin T Myoglobin and CK-MB cardiac disease

The Troponin T/Myoglobin/CK-MB(3 in 1)Test Cassette (Whole Blood/Serum/Plasma) is intended for in vitro quantitative determination of human cardiac Troponin T, Myoglobin and CK-MB in whole blood, serum or plasma as an aid in the diagnosis of Myocardial Infarction (MI).

【SUMMARY】

Myoglobin (MYO), Creatine Kinase MB (CK-MB) and cardiac Troponin T (cTnT) are proteins released into the bloodstream after cardiac injury. Myoglobin is a heme-protein normally found in skeletal and cardiac muscle with a molecular weight of 17.8 kDa.¹ When muscle cells are damaged, Myoglobin is released into the blood rapidly due to its relatively small size. The level of Myoglobin increases measurably above baseline within 2-4 hours post-infarct, peaking at 9-12 hours, and returning to baseline within 24-36 hours.^2,3 CK-MB is an enzyme also present in the cardiac muscle, with a molecular weight of 87.0 kDa.⁴ Creatine Kinase is a dimeric molecule formed from two subunits designated as “M” and “B”, which combine to form three different isoenzymes, CK-MM, CK-BB and CK-MB. CK-MB is the isoenzyme of Creatine Kinase most involved in the metabolism of cardiac muscle tissue.⁵ The release of CK-MB into the blood following an MI can be detected within 3-8 hours after the onset of symptoms. It peaks within 9 to 30 hours, and returns to baseline levels within 48 to 72 hours.⁶ Cardiac Troponin T(cTnT) is a structurally bound protein found in striated muscle cells with a molecular weight of 37kD.⁷ Troponin T is part of a three subunit complex comprising of Troponin I and Troponin C. Along with tropomyosin, this structural complex forms the main component that regulates the calcium sensitive ATPase activity of actomyosin in striated skeletal and cardiac muscle.⁸ After acute myocardial infarction (AMI), serum cTnT levels are elevated 2 to 8 hours after onset, peak in 12-24 hours and can persist for up to 14 days.⁹ Thus providing for a longer window of detection for cardiac injury.

The combined detection of three myocardial injury markers including CK-MB, cTnT and myoglobin has better sensitivity and specificity, which makes the diagnosis more accurate. To achieve early diagnosis, early treatment, reduce missed diagnosis and timely risk stratification of patients.

The Troponin T/Myoglobin/CK-MB(3 in 1)Test Cassette (Whole Blood/Serum/Plasma) is a simple test that utilizes a combination of antibody coated particles and capture reagent to detect cardiac Troponin T, Myoglobin and CK-MB in whole blood, serum or plasma.

【PRINCIPLE】

The Troponin T/Myoglobin/CK-MB(3 in 1)Test Cassette (Whole Blood/Serum/Plasma) detects cardiac Troponin T, Myoglobin and CK-MB based on Fluorescence Immunoassay. The sample moves through the strip from sample pad to absorbent pad. If the specimen contains cTnT, Myoglobin or CK-MB, it attaches to the fluorescent microspheres-conjugated specific antibodies. Then the complex will be captured by the capture antibodies coated on the nitrocellulose membrane (Test line). The concentration of cTnT, Myoglobin and CK-MB in the sample correlates with the fluorescence signal intensity captured on the Test line. According to the fluorescence intensity of the test and product standard curve, the concentration of cTnT, Myoglobin and CK-MB in the sample can be calculated by FIATEST^TM Analyzer to show cTnT, Myoglobin and CK-MB concentration in specimen.

【REAGENTS】

The test kit includes cTnT antibody coated fluorophores, Myoglobin antibody coated fluorophores, CK-MB antibody conjugated fluorophores and capture reagents coated on the membrane.

【PRECAUTIONS】

• For professional in vitro diagnostic use only.
• Do not use after the expiration date indicated on the package. Do not use the test

if the foil pouch is damaged. Do not reuse.

• Avoid cross-contamination of specimens by using a new specimen collection container for each specimen obtained.
• Do not eat, drink or smoke in the area where the specimens and tests are handled. Handle all specimens as if they contain infectious agents. Observe established precautions against microbiological hazards throughout the procedure and follow standard procedures for proper disposal of specimens. Wear protective clothing such as laboratory coats, disposable gloves and eye protection when specimens are assayed.
• Do not interchange or mix reagents from different lots.
• Humidity and temperature can adversely affect results.
• Used testing materials should be discarded in accordance with local regulations.
• Read the entire procedure carefully prior to any testing.
• The FIATEST^TM Troponin T/Myoglobin/CK-MB(3 in 1)Test Cassette should only be used with the FIATEST^TM Analyzer by approved medical professionals.

【STORAGE AND STABILITY】

• The test should be stored at 4-30 °C until the expiry date printed on the sealed pouch.
• The test must remain in the sealed pouch until use.
• Do not freeze.
• Care should be taken to protect the components of the kit from contamination.
• Do not use if there is evidence of microbial contamination or precipitation. Biological contamination of dispensing equipment, containers or reagents can lead to false results.

【SPECIMEN COLLECTION AND PREPARATION】

Preparation

• Before performing the test, please make sure that all components are brought to room temperature(15-30 ℃). Cold buffer solution or moisture condensation on the membrane can lead to invalid test results.
• Take a tube with buffer solution out of the kit. Document patients name or ID on it.

Sample Handling

• Collect the specimen according to standard procedures.
• Do not leave specimens at room temperature for prolonged periods. Serum and plasma specimens may be stored at 2-8 °C for up to 1 day, for long term storage, specimens should be kept below -20 °C. Whole blood collected by venipuncture should be stored at 2-8 °C if the test is to be used within 1 day of collection. Do not freeze whole blood specimens. Whole blood collected by finger stick should be tested immediately.
• Bring specimens to room temperature prior to testing. Frozen specimens must be completely thawed and mixed well prior to testing. Avoid repeated freezing and thawing of specimens.
• EDTA K2, Heparin sodium, Citrate sodium and Oxalate potassium can be used as the anticoagulant tube for collecting the blood specimen.

Sample Dilution

• The specimen (75 μL of serum/plasma/whole blood) can be added directly with the micro pipette into the buffer.
• Close the tube and shake the sample by hand for approximately 10 seconds so sample and dilution buffer mix well.
• Let the diluted sample homogenize for approximately 1 minute.
• It is best to place the diluted sample on an ice pack and leave the sample at room temperature for no more than 2 hours.

【MATERIALS】

Materials Provided

• Test Cassettes • Specimen Collection Tubes with Buffer

• ID Card • Package Insert

Materials Required But Not Provided

• Timer • Centrifuge • FIATEST^TM Fluorescence Immunoassay Analyzer

• Pipette

【DIRECTIONS FOR USE】

Refer to FIATEST^TM Fluorescence Immunoassay Analyzer Operation Manual for the complete instructions on use of the Test. The test should be conducted at room temperature.

Allow the test, specimen, buffer and/or controls to reach room temperature (15-30 °C) prior to testing.

• Turn on the Analyzer power. Then according to the need, select “standard test” or “Quick test” mode.
• Take out the ID card and insert it into the Analyzer port.
• Serum/plasma: Transfer 75 μL serum/plasma into the buffer tube, mix the specimen and the buffer well.

Whole blood: Transfer 75 μL whole blood into the buffer tube with pipette; mix the specimen and the buffer well.

• Add diluted specimen with a Pipette: Pipette 75 μL diluted specimen into the sample well. Start the timer at the same time.
• There are two test modes for FIATEST^TM Fluorescence Immunoassay Analyzer, Standard Test mode and Quick Test mode. Please refer to the user manual of FIATEST^TM Fluorescence Immunoassay Analyzer for details.

“Quick test” mode: Insert the test cassette into the Analyzer at 15 minutes after sample application, click “QUICK TEST”, fill the test information and click "NEW TEST" immediately. The Analyzer will automatically give the test result after a few seconds.

“Standard test” mode: Insert the test cassette into the Analyzer immediately after sample application, click “STANDARD TEST”, fill the test information and click "NEW TEST" at the same time, the Analyzer will automatically count down 15 minutes. After the countdown, the Analyzer will give the result at once.

【INTERPRETATION OF RESULTS】

Results read by FIATEST^TM Fluorescence Immunoassay Analyzer.

The result of tests for cTnT, Myoglobin and CK-MB is calculated by FIATEST^TM Fluorescence Immunoassay Analyzer and display the result on the screen. For additional information, please refer to the user manual of FIATEST^TM Fluorescence Immunoassay Analyzer.

Linearity range of cTnT Test is 0.1-40 ng/mL.

Linearity range of Myoglobin Test is 5-200 ng/mL.

Linearity range of CK-MB is 0.2-75 ng/mL.