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Cardiac muscle troponin I detection reagent kit and its preparation method

A technique for cardiac troponin and kit, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of slow cardiac troponin I, low sensitivity, complicated operation, etc., and achieves strong specificity, high sensitivity, Simple to use effects

Active Publication Date: 2008-01-16
SHANGHAI CHEMTRON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The purpose of the present invention is to overcome the shortcomings of the prior art, such as enzyme-linked immunoassay and luminescence immunoassay, which detect cardiac troponin I at a slow speed, complicated operation and low sensitivity, and provide a fast, convenient and sensitive non-toxic Cardiac troponin I detection kit relying on specialized equipment

Method used

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  • Cardiac muscle troponin I detection reagent kit and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Preparation of monoclonal antibody to cardiac troponin I.

[0038] (1) Separation and purification of cardiac troponin I

[0039] a) Take the myocardium of a patient who died of heart disease, soak it in 0.2M acetic acid-sodium acetate buffer solution with a pH value of 5.6, homogenize it with a high-speed mixer, centrifuge at 7000-10000rpm for 30 minutes, and take the supernatant and store it at 68°C Heat treatment for 3 minutes, centrifuge at 10,000rpm for 30 minutes to take the supernatant, and put it on a DEAE cellulose chromatography column for ion exchange chromatography, the eluent is pH5.6, 0.1M acetic acid-sodium acetate buffer, and the elution flow rate is 60ml / h , eluted with 3 column volumes; collected the eluent, and put it on a SephadexG75 gel filtration chromatography column, the eluent was pH7.0-7.4, 0.2M phosphate buffer, the elution flow rate was 20ml / h, and the effluent was collected first peak.

[0040] b) The molecular weight and purity of the pur...

Embodiment 2

[0047] Preparation of detection kits for diagnosing myocardial infarction

[0048] (1) Preparation of colloidal gold: use trisodium citrate to reduce chloroauric acid to obtain colloidal gold with a particle size of 40-60nm

[0049] a) Prepare 0.01% HauCl aqueous solution and 1.0% trisodium citrate aqueous solution respectively;

[0050] b) Put 1000ml of 0.01% HauCl solution in a beaker, and heat it to boiling on an electric stove. Add 6.5 ml of 1.0% trisodium citrate aqueous solution, and stir while adding. And continue heating and boiling for 10-15 minutes until the solution is transparent red. Cool to room temperature, add purified water to restore the original volume.

[0051] (2) Preparation of colloidal gold-labeled anti-cTnI monoclonal antibody:

[0052] a) Dilute the anti-cTnI monoclonal antibody obtained in Example 1 with pH 7.2, 0.1M phosphate buffer to a protein concentration of 1.2 mg / ml;

[0053] b) adding the diluted anti-cTnI monoclonal antibody solution to...

Embodiment 3

[0059] Clinical use of cardiac troponin I detection kit:

[0060] (1) Take 45 parts of the test kit prepared in the above-mentioned embodiment 2, select 20 blood samples of myocardial infarction patients and 25 normal human blood samples, and add two to three drops of blood samples to the sample holes of the kit respectively one-to-one. All 45 kits developed color within 10-15 minutes after adding the sample. Reach the expected purpose of displaying the results visible to the naked eye within 15 minutes. Results The test results of 20 patients with acute infarction were all positive, and the test results of 25 normal subjects were all negative. There was no uncolored kit, which indicated that the kit was accurate and rapid.

[0061] (2) Interpretation of results: Interpret the results within 10 to 15 minutes after adding the sample. If a red color band appears at the detection line and the quality control line respectively, it means that there is cTnI in the tested sample, a...

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Abstract

The invention relates to a reagent kit and the preparation method thereof which are used to detect cardiac muscle troponin I; the reagent kit provided by the invention comprises a reagent strip; the reagent strip contains cardiac muscle troponin I monoclonal antibody labeled by colloid gold, unlabeled cardiac muscle troponin I monoclonal antibody and the second antibody of anti-myocardial troponin I monoclonal antibody; the invention also provides the preparation method of the reagent kit of detection. The reagent kit of detection can detect cardiac muscle troponin I in blood sample drawn from human body swiftly, conveniently and delicately; the cardiac muscle troponin I can be used to diagnose myocardial infarction.

Description

[0001] technical field [0002] The invention relates to a test kit for detection, in particular to a detection kit for diagnosing myocardial infarction by utilizing cardiac troponin-related immune reaction immunochromatographic color development and a preparation method thereof. Background technique [0003] As the number one killer that endangers human life and health—the incidence of cardiovascular and cerebrovascular diseases is increasing year by year. Every year, there are 450 million patients with symptoms of acute myocardial infarction (AMI) in the emergency department in the United States, and 400 million patients are identified as AMI, there are 270 million patients who are not AMI, and about 5 million patients are diagnosed as non-AMI, but 2-3% of them really have AMI and because the symptoms of AMI are various, there are many complications in clinical practice. Misdiagnosis and missed diagnosis. Some existing myocardial injuries are ignored by doctors and patients ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/549G01N33/531
Inventor 张国华
Owner SHANGHAI CHEMTRON BIOTECH
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