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Troponin I detection reagent and preparation method thereof

A detection reagent and troponin technology are applied in the field of troponin I detection reagent and preparation thereof, homogeneous sol particle type cTnI immunoassay reagent and preparation thereof, and can solve the problems of low initial sensitivity, high specificity and the like, Achieve high analytical sensitivity and high specificity

Inactive Publication Date: 2015-02-04
中国人民解放军第三0五医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

cTnI shows lower initial sensitivity and higher specificity than cTnT

Method used

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  • Troponin I detection reagent and preparation method thereof
  • Troponin I detection reagent and preparation method thereof
  • Troponin I detection reagent and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0031] The preparation method of described Troponin I detection reagent is as follows:

[0032] 1), adding coagulant, BSA and preservative to the buffer to prepare reagent one;

[0033] 2), heating and boiling chloroauric acid and trisodium citrate according to the mass ratio of 1:1 to obtain a colloidal gold solution; adding the cTnI antibody to the colloidal gold solution, and adding a stabilizer, a buffer, and a preservative to obtain a reagent 2. Among them, when configuring chloroauric acid and trisodium citrate solution, use 0.2 μm filter membrane to filter;

[0034] 3), the first reagent and the second reagent are used together according to the volume ratio of 5:1 to obtain the troponin I detection reagent.

Embodiment 1

[0037] Preparation of colloidal gold particles (all the following operations need to be completed in a high-cleanliness dust-free space)

[0038] 1) Preparations: All glass containers used are first washed with detergent and rinsed with running water, then soaked with silicification reagent overnight, and the inner surface of all gold-making utensils is silicified, and then rinsed with distilled water for use;

[0039] 2) Prepare 1% (w / v) chloroauric acid and 1% (w / v) sodium citrate solution, and filter with 0.2 μm membrane;

[0040] 3) Put 1 magnetic stirring bar in the cleaned 1000ml round bottom flask, and add 1000ml ultrapure water;

[0041] 4) Add a certain volume concentration of 1% (w / v) chloroauric acid, stir at a high speed around 600rpm, and heat until the solution boils, then quickly add a certain volume concentration of 1% sodium citrate solution to the flask, keep heating And vigorously stirred for 10min, the color of the solution turned black at first, and then ...

Embodiment 2

[0045] Preparation of reagent one

[0046] The formula of reagent 1 is as follows: Tris-HCl buffer, 0.9% NaCl (w / v), 2% BSA (w / v), 0.1% NaN3 (w / v), 0.05% Tween20 (w / v), according to The concentration of the quality control buffer of Tris, the pH of the buffer is adjusted according to the amount of HCl used.

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Abstract

The invention relates to a troponin I detection reagent and a preparation method thereof. The troponin I detection reagent is prepared from a first reagent and a second reagent in the volume ratio of 5: 1, wherein the first reagent contains 20-50mmol / L of buffer solution, 2%w / v of coagulant, 2%w / v of BSA and 0.1%-0.5%w / v of preservative; the second reagent contains 0.1%-0.5%w / v of colloid gold particles marked with anti-human cTnI antibodies, 2%-5%w / v of stabilizer, 20-50mmol / L of buffer solution and 0.1%-0.5%w / v of preservative; the diameter of the colloid gold particles marked with the anti-human cTnI antibodies ranges from 60nm to 90nm. The troponin I detection reagent has the advantages of being good in operability, high in analytical sensitivity, high in linearity, and quick, simple and convenient.

Description

【Technical field】 [0001] The invention relates to a troponin I detection reagent and a preparation method thereof, in particular to a homogeneous sol particle-type cTnI immunoassay reagent for detecting cTnI in serum or plasma and a preparation method thereof, belonging to the in vitro diagnosis of medical equipment field. 【Background technique】 [0002] Troponin I has a molecular weight of 23,000 and is an actin inhibitory subunit. It has three subtypes: fast skeletal muscle subtype, slow skeletal muscle subtype and cardiac subtype. The three TnI isoforms are derived from three different genes. The cardiac isoform (cTnI) relative to the two skeletal muscle isoforms is about 40% different. The released form of cTnI after AMI (free form or complex with other troponins, oxidized form or reduced form) has so far been unknown. Not entirely clear. [0003] Diagnosis of Myocardial Injury Among the many clinical biochemical indicators for the diagnosis of acute myocardial infarc...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6887
Inventor 王会中
Owner 中国人民解放军第三0五医院
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