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Detection kit for cardiac troponin I and preparation method thereof

A technique for cardiac troponin and detection kit, which is applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of low sensitivity, long detection period, and small detection concentration range, so as to improve sensitivity and ensure accuracy. The effect of performance and precision, efficient detection

Active Publication Date: 2019-01-18
SICHUAN MACCURA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The ELISA method has a long detection cycle; the chemiluminescence method is expensive and requires special instruments and professionals to operate, and is only suitable for use in central laboratories. The detection cycle is long and cannot meet the needs of clinical rapid detection, and it is not suitable for small and medium-sized hospitals, especially small and medium-sized hospitals. Community hospitals and township hospitals without a central laboratory; and fluorescence immunochromatography can achieve rapid quantitative detection of emergency samples
[0004] At present, there are problems such as low sensitivity and small detection concentration range in the immunochromatographic assay method of cardiac troponin I in the prior art. Therefore, it is of great significance to develop efficient cardiac troponin detection reagents and detection methods

Method used

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  • Detection kit for cardiac troponin I and preparation method thereof
  • Detection kit for cardiac troponin I and preparation method thereof
  • Detection kit for cardiac troponin I and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] The preparation of embodiment 1 detection kit

[0061] 1. Preparation of the sample pad: use glass fiber membrane as the sample pad, place the glass cellulose membrane on a white PVC board, prepare 0.1% Tween-20 solution as the sample pad treatment solution, and use 45 μL / cm 2 Sprinkle the sample pad treatment solution evenly on the glass cellulose membrane, spread evenly on the front and back of the nitrocellulose membrane until there are no white spots, and then place it on the screen window to balance at room temperature for 60min±15min. , dried at 37°C for 30-72 hours;

[0062] 2. Preparation of reaction solution:

[0063] (1) Polypropylene fluorescent microspheres (using Eu as the fluorescent substance, the particle size of the fluorescent microspheres is 200nm, and the fluorescent microspheres contain carboxyl modification) diluted 10 times with 20mM MES (2-morpholineethanesulfonic acid) (pH 6.0) , ultrasonic 1min;

[0064] (2) Prepare 5mg / mL EDC (1-(3-dimethyl...

Embodiment 2

[0079] The sensitivity analysis that embodiment 2 kit detects

[0080] 1. Preparation of cTnI detection standard

[0081] Use normal human serum as a diluent to prepare cTnI antigen standard products with concentrations of 0 ng / mL and 50 ng / mL respectively; mix the two concentrations of cTnI standard product solutions in sequence according to a certain concentration, and prepare 0-50 ng / mL respectively Concentration gradient of cTnI detection standard.

[0082] 2. Detection reaction

[0083] Dilute the reaction solution in the kit prepared in Example 1 by 10 times with the diluent, add 60 μL of the detection standard of each concentration obtained in step 1, and mix well for 5 seconds to 10 seconds.

[0084] Pipette 90 μL of the mixture of the detection standard and the reaction solution at each concentration above, and add it dropwise to the sample hole on the sample pad of the test paper card. Be careful not to inhale air bubbles when sampling, and place it at room tempera...

Embodiment 3

[0090] Accuracy and precision analysis of embodiment 3 kit detection

[0091] The kit prepared in Example 1 was used to detect cTnI in 3 clinical serum samples. According to the content of cTnI in the three samples, they are divided into three groups: low value, middle value and high value, among which the clinical measurement value of the low value fixed value sample is 0.15ng / mL, and the clinical measurement value of the middle value fixed value sample is 0.57ng / mL , the clinical measurement value of high-value samples was l.89ng / mL. Each of the three fixed-value samples was tested 10 times, and the test results are shown in Table 2. The results show that the test kit prepared in Example 1 is used to detect 3 samples of fixed value, and the test results are very close to the clinical measurement values ​​(the difference between the average value of 10 tests and the clinical measurement values ​​is only about 0.01 ng / mL), It shows that the cTnI detection kit of the present ...

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Abstract

The invention relates to a detection kit for cardiac troponin I and a preparation method thereof. The detection kit for cardiac troponin I disclosed by the invention comprises an detection antibody and tracer-marked capture antibodies, wherein the capture antibodies are respectively two monoclonal antibodies combining different loci of the cardiac troponin I, and the detection antibody is a polyclonal antibody of the cardiac troponin I. The detection kit for cardiac troponin I and the preparation method thereof provided by the invention obviously improve the detection sensitivity and linear range, ensure accuracy and degree of precision at the same time, and can be used for efficient detection of cardiac troponin I by screening proper capture antibodies, detection antibody and combinationthereof detected by immunochromatography of the cardiac troponin I, and optimizing diluent components of the kit synchronously.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a cardiac troponin I detection kit and a preparation method thereof. Background technique [0002] Cardiac Troponin I (cTnI) is a subunit of cardiac troponin, present on the thin filaments of cardiac myofibrils, and is a unique structural protein of cardiomyocytes. Under normal circumstances, the content of cTnI in the blood is very low. When myocardial injury occurs, cTnI is rapidly released into the blood, resulting in an increase in the content of cTnI in the blood. After 3 to 5 hours of cardiomyocyte injury, cTnI in peripheral blood rises, reaches the peak at 12 to 24 hours, and lasts for a long time, up to 5 to 8 days, and basically returns to normal in about 10 days. Cardiac troponin I can be used as an auxiliary clinical diagnosis of acute myocardial infarction (AMI), an indicator of short-term risk classification, prognosis assessment and dynamic monitoring of acute c...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/54313G01N33/6887
Inventor 姚凯成刘东泽赵书阁
Owner SICHUAN MACCURA BIOTECH CO LTD
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