Cardiac troponin I (cTnI)

Cardiac Troponin I (cTnI) Assay Kit

Instructions for Use (Magnetic Microparticle Chemiluminescence Method)

Instruction of Cardiac Troponin Ⅰ（cTnⅠ）Detection Kit（Magnetic Solid Phase Chemiluminescent Immunoassay）

【Product Name】

Generic Name: Cardiac Troponin I (cTnI) Assay Kit (Magnetic Microparticle Chemiluminescence Method)

English Name: Cardiac Troponin Ⅰ（cTnⅠ）Detection Kit（Magnetic Solid Phase Chemiluminescent Immunoassay)

【Packaging Specifications】

50 tests/box, 100 tests/box, 200 tests/box.

Calibrator: 6 × 1 bottle (optional), 2 × 1 bottle (optional)

Quality Control: 2 × 1 bottle (optional)

【Intended Use】

The Cardiac Troponin I (cTnI) Assay Kit (Magnetic Microparticle Chemiluminescence Method) (hereinafter referred to as this kit) is used for the in-vitro quantitative detection of Cardiac Troponin I (cTnI) in human serum.

Used for the diagnosis of acute myocardial infarction (AMI), post-operative myocardial injury, and chemotherapy cardiotoxicity, as well as other myocardial injuries related to diseases. cTnI has high myocardial tissue specificity. In the blood, it mainly exists as a cTnI complex, with less free form, mostly in a molecular form. Under normal circumstances, the serum cTnI concentration is extremely low. When the myocardium is damaged, the myocardial cell membrane is also damaged, and cTnI is rapidly released into the blood. cTnI reaches various myocardial interstitiums through the blood, and can be detected in the blood 4-6 hours after the onset of the disease. Relevant studies have shown that the cTnI concentration is closely related to the prognosis of AMI patients, especially the low-value detection area of cTnI can provide rapid early diagnosis, which has important clinical significance. Due to the high myocardial cell specificity and high sensitivity of cTnI, cTnI has been widely accepted clinically, not only as the "gold standard" for diagnosing acute myocardial infarction, but also as the most suitable marker for monitoring the condition, observing the efficacy, and evaluating the prognosis of myocardial diseases. Common laboratory diagnostic methods in clinical practice include immunochromatography and latex immunoturbidimetry.

【Test Principle】

This kit uses a direct sandwich method: Biotin-labeled cTnI antibody, acridinium ester (AE)-labeled cTnI antibody, and cTnI in the sample, calibrator, or quality control bind to form a "sandwich" complex. Then, streptavidin-coated magnetic microparticles are added. Through the specific binding of streptavidin and biotin, the antigen-antibody complex is connected to the magnetic particles. Under the action of an external magnetic field, the complex formed by the immunoreaction is separated from other unbound substances. After washing the complex, a pre-excitation solution and an excitation solution are added. Acridinium ester forms an unstable excited intermediate under the action of the pre-excitation solution and the excitation solution. When the excited intermediate returns to the ground state, it emits photons, forming a chemiluminescent reaction, which can be detected using a chemiluminescence instrument. Within the detection range, the chemiluminescence intensity is proportional to the cTnI concentration in the sample. The cTnI concentration in the sample can be calculated using a four-parameter Logistic equation.

【Main Components】

1. Product Composition

cTnI magnetic microparticle reagent: 1 bottle, streptavidin-coated magnetic microparticles;

cTnI reagent 1: 1 bottle, biotin-labeled cTnI antibody conjugate;

cTnI reagent 2: 1 bottle, acridinium ester (AE)-labeled cTnI antibody conjugate;

Calibrator (lyophilized powder): 6 bottles or 2 bottles (optional), containing BSA buffer with different amounts of cTnI antigen added. The calibrator value is obtained by scanning the calibrator QR code. After dissolving at room temperature, it is detected;

Quality control (lyophilized powder): 1 bottle each of high and low points (optional), containing BSA buffer with different amounts of cTnI antigen added. The target value range of the quality control is obtained by scanning the quality control QR code. After dissolving at room temperature, it is detected.

Note ①: Reagents from different batches of kits are prohibited from being used interchangeably.

Note ②: Six calibrator points or two calibrator points are optional. If calibrators and quality controls have been provided to the user, the kit does not need to be reconfigured.

2. Matching Reagents

The following matching reagents are not included in this product but are required for detection:

Pre-excitation solution and excitation solution: Excite acridinium ester to catalyze the generation of photons, so that the chemiluminescence intensity can be detected using a chemiluminescence immunoassay instrument.

Washing solution: Diluted for washing the reaction system.

Source: Provided by our company.

Note: To ensure the accuracy of the test results, the manufacturer of the matching reagents must not be changed arbitrarily during the test.

3. Calibrator Traceability

Calibrators are traceable to the enterprise's selected measurement procedure.

【Storage Conditions and Shelf Life】

The reagents are stable for 15 months at 2-8℃, protected from light, and should not be frozen.

After opening the liquid reagent, it is stable for 30 days at 2-8℃, protected from light.

【Production Date】

See product label.

【Applicable Instruments】

Cosmome SMART500S, 6500 or Tuochuang Medical TC-300 fully automatic chemiluminescence immunoassay instrument

【Sample Requirements】

1. This kit is suitable for serum and whole blood samples.

2. After blood collection, serum should be separated immediately for analysis to avoid hemolysis. Testing should be completed within 24 hours at room temperature, stable for 3 days at 2-8℃, and stable for 30 days at -20℃. Avoid repeated freezing and thawing.

3. Frozen and refrigerated samples should be returned to room temperature and mixed thoroughly before use.

4. Avoid using samples with severe hemolysis or high concentrations of lipemia.

5. The sample volume should be no less than 200μL.

【Test Method】

Note: Please read the instructions carefully before use. Working conditions: Room temperature (15～35℃); Relative humidity ≤80%.

1. Pre-test preparation

1.1 Before the test, all reagents should be placed at room temperature, and the instrument should be preheated for at least 30 minutes;

1.2 According to the system operating instructions, perform the loading of reaction tubes (cups), the addition or replacement of matching reagents, waste liquid, waste tube cleaning, and liquid path filling operations.

2. Reagent and sample loading

2.1 Before reagent loading, place the reagents to be used on the mixing equipment for thorough mixing. Visually inspect the reagent solution components; they should be clear, free of foreign matter, precipitates, and flocculent matter. Magnetic microparticle reagents should be a uniform suspension without obvious aggregation;

2.2 According to the system operating instructions, scan the reagent barcodes to complete the loading of magnetic microparticle reagents and anti-reagents;

2.3 After mixing the calibrators and quality control samples, transfer them to the instrument's dedicated reaction cups, load them into the instrument's sample positions, and load the centrifuged serum samples directly into the instrument's sample positions after scanning.

3. Detection steps

For optimal detection performance, please follow the relevant instructions in this instruction manual and operate according to the operating manual of the fully automatic chemiluminescence analyzer.

Note: The detection reaction process and related parameters have been predefined in the instrument operating software.

4. Calibration curve and calibration

The calibration curve is obtained by scanning the main curve card to generate it directly, or by directly detecting six calibrator points to generate a fit, and the system also supports obtaining a working calibration curve using a two-point calibration method.

Note ①: Due to inter-instrument differences and system differences caused by different operators, operating environments, and matching general reagents, the six-point direct calibration method is recommended to obtain the working calibration curve.

Note ②: Due to reagent activity drift and changes in general reagent batches, the calibrated working curve needs to be recalibrated after a certain period of use. Recalibration should be performed when the following situations occur:

l After 1 month (28 days) of using the same batch number of reagents;

l Using a new batch number of reagents, the whole box or pre-activation liquid and activation liquid;

l When the quality control values are outside the quality control range;

l After each maintenance of the analyzer.

5. Result output

The analyzer automatically calculates the cTnI concentration of each sample using a working curve obtained by two-point calibration from the standard curve, and the results are expressed in ng/mL.

6. Quality control

Two levels of quality control samples should be determined daily along with sample testing, treating the quality control samples as patient samples;

Note: It is recommended to use at least two levels of commercial quality control products. If the test results are within the system's acceptable quality control range or within your specified range (determined by an appropriate internal laboratory quality control program), the test results are satisfactory; otherwise, it indicates that the test results are unreliable and a test report should not be issued.

【Reference Range】

Normal range of this kit: ≤0.04ng/mL.

Note: The measured cTnI levels will vary depending on the region, individual, and method used for testing. Therefore, we recommend that each laboratory establish its own normal range. A diagnosis should not be made solely based on the cTnI value obtained by this method; the results should be analyzed in conjunction with other clinical data, including the patient's specific condition and treatment status.

【Interpretation of Test Results】

1. Due to methodological or antibody specificity reasons, testing the same sample using reagents from different manufacturers may yield different results. Therefore, results obtained using different reagents should not be directly compared to avoid erroneous medical interpretations.

2. Test results exceeding the measurement range of the kit are calculated results obtained by extrapolating the calibrator curve. When reporting such results, please pay special attention. If an accurate value is desired, dilute the sample appropriately before testing; the maximum dilution factor is 10 times.

3. Quality control products can serve as a reference for the reliability of the results of the current experiment; their measured values should be within the allowable range specified on the quality control sheet for this batch of products. Test results should be judged comprehensively based on the reference range and other clinical factors and results. When the test result is close to the upper limit of the reference range, consider performing a confirmatory test on the sample.

【Limitations of the Test Method】

1. Severe hemolysis (hemoglobin ≥500mg/dL), lipemia (triglycerides ≥1500mg/dL), jaundice (bilirubin ≥20mg/dL), and contaminated samples may affect the test results and should be avoided;

2. The test results of this kit are for clinical reference only and cannot be used alone as the basis for confirming or excluding cases. To achieve diagnostic purposes, this test result should be used in conjunction with clinical examinations, medical history, and other examinations.

3. This product can be used for the determination of human serum samples, and the reliability of its use for the determination of cTnI concentration in other body fluids has not been fully confirmed.

【Product Performance Indicators】

1. Reagent performance indicators

1.1 Accuracy: Recovery rate P is 85%-115%.

1.2 Precision: Intra-batch precision ≤8%, inter-batch precision ≤10%.

1.3 Blank limit: Should not exceed 0.04 ng/mL.

1.4 Linearity: Linear range (0.04～50.0) ng/mL, correlation coefficient r≥0.9900.

1.5 Specificity: When the concentration of cardiac troponin C (cTnC), cardiac troponin T (cTnT), and skeletal muscle troponin I (sTnI) is 1000 ng/mL, the cross-reactivity rate should not exceed 1%.

2. Calibrator Performance Indicators

2.1 Accuracy: Recovery rate P is between 85%-115%.

2.2 Uniformity: Inter-bottle difference (CV) ≤8%.

3. Quality Control Product Performance Indicators

3.1 Expected results: Test values should be within ±15% of the target value of the quality control product.

3.2 Uniformity: Inter-bottle difference (CV) ≤8%.

【Precautions】

1. This reagent is for in vitro diagnostic use only. Reagents from different batches are prohibited from being used interchangeably. Do not use expired reagents.

2. Some components of this kit contain bovine-derived materials, but these raw materials are not from countries or regions with bovine spongiform encephalopathy (BSE) outbreaks, and their production is not carried out in countries or regions with BSE outbreaks. If it is bovine serum, it will be filtered and sterilized after purchase before being used in kit production. Therefore, the kit does not pose a risk of transmitting BSE; some components of the kit contain porcine-derived materials, but these raw materials are not from countries or regions with foot-and-mouth disease outbreaks, and their production is not carried out in countries or regions with foot-and-mouth disease outbreaks. If it is porcine serum, it will be filtered and sterilized after purchase before being used in kit production. Therefore, the kit does not pose a risk of transmitting foot-and-mouth disease; this reagent does not contain human serum, but should still be considered a potential biohazard.

3. This reagent contains preservatives. Avoid contact with skin. If skin accidentally comes into contact with the reagent, rinse thoroughly with plenty of water.

4. All reagents should be stored at 2～8℃, avoid freezing at -20℃, and equilibrate to room temperature before use.

5. The magnetic microparticle reagent should be thoroughly mixed before use, and avoid strong shaking.

【Explanation of Labels】

Store at 2-8℃ Avoid direct sunlight Store upright

In vitro diagnostic reagent Production date Expiration date

Product batch number See instructions for details

【References】

1. Sandoval Y and Thygesen K. Myocardial Infarction Type 2 and Myocardial Injury. Clin Chem. 2017, 63(1):101-107.

2. Thygesen K, Alpert JS, White HD, et al．Universal definition of myocardial infarction．Circulation. 2007, 116 ( 22) :2634-2653．

3. Liu Xiaoge, Pan Xiaoyan, Liu Aisheng, et al. Clinical value of combined detection of h-FABP, Mb, cTnI and CK-MB in early diagnosis of myocardial injury in children with hand, foot and mouth disease. Chinese Medical Equipment. 2016, 13(10):89-92.

4. Jia Xiaobing. Clinical significance of quantitative detection of cardiac troponin I in acute myocardial infarction. China Prescription Drugs. 2015, 000(002):115-116.

【Basic Information】

Registered person/manufacturer: Zhejiang Tuochuang Medical Technology Co., Ltd

Address: No. 1 Building, Qianren Plan Industrial Park, No. 78, Zhancheng Avenue, Zhuji City, Zhejiang Province

Production address: No. 1 Building, Qianren Plan Industrial Park, No. 78, Zhancheng Avenue, Zhuji City, Zhejiang Province

After-sales service unit: Zhejiang Tuochuang Medical Technology Co., Ltd

Production license number:

Contact number: 0575-87178692

Fax:

Post code: 311800

Website: www.zjtcyl.com

【Medical Device Registration Certificate Number/Product Technical Requirements Number】

【Date of Approval and Revision of Instructions for Use】

Approval date:

Revision date: