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Kit for quantitative detection of H-FABP content by using proximity ligation assay technology, preparation method and use method thereof

A technology of H-FABP and connection technology, applied in the field of adjacent connection technology, can solve the problems of low sensitivity, poor repeatability and difficult quantification of immunochromatography

Inactive Publication Date: 2018-08-10
NANJING TZONE BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have corresponding shortcomings. The sensitivity of immunochromatography is low, the reagents are unstable, the repeatability is poor, and it is difficult to quantify; the stability of liquid latex reagents in immunoturbidimetry is not good, and the accuracy of detection results is not good.

Method used

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  • Kit for quantitative detection of H-FABP content by using proximity ligation assay technology, preparation method and use method thereof
  • Kit for quantitative detection of H-FABP content by using proximity ligation assay technology, preparation method and use method thereof
  • Kit for quantitative detection of H-FABP content by using proximity ligation assay technology, preparation method and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Preparation of H-FABP probe A:

[0026] 1) Take 0.1mg of anti-H-FABP antibody and incubate with DBCO-sulfo-NHS reagent for 5-30min;

[0027] 2) After the reaction is over, add 10 μL of 0.5mol / L Tris-HCl and incubate for 5-10 minutes to terminate the reaction;

[0028] 3) Add the above-mentioned modified antibody to oligodeoxynucleotide a, and incubate overnight at 4°C to obtain H-FABP probe A.

Embodiment 2

[0030] Preparation of H-FABP probe B:

[0031] 1) Take 0.1mg of anti-H-FABP antibody and incubate with DBCO-sulfo-NHS reagent for 5-30min;

[0032] 2) After the reaction is over, add 10 μL of 0.5mol / L Tris-HCl and incubate for 5-10 minutes to terminate the reaction;

[0033] 3) Add the above-mentioned modified antibody to oligodeoxynucleotide b, and incubate overnight at 4°C to obtain H-FABP probe B.

Embodiment 3

[0035] The main components of the kit:

[0036] 1) H-FABP probe A;

[0037] 2) H-FABP probe B;

[0038] 3) connection buffer;

[0039] 4) qPCR reaction solution.

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Abstract

The present invention provides a kit for quantitative detection of H-FABP content by using an proximity ligation assay technology, wherein the kit comprises an H-FABP probe A, an H-FABP probe B, a ligation buffer liquid, and a qPCR reaction solution. The present invention further discloses a preparation method of the kit, wherein the preparation method comprises: preparation of an H-FABP probe A,and preparation of an H-FABP probe B. The invention further discloses a use method of the kit. According to the present invention, the proximity ligation assay technology is firstly applied in the quantitative detection of H-FABP content; and compared to the method in the prior art, the kit of the present invention has advantages of high sensitivity, high specificity, high throughput and the like,can improve the accuracy of the early diagnosis of acute coronary syndrome, and has great market value.

Description

technical field [0001] The invention relates to an adjacent junction technique used for in vitro immunodiagnosis to detect the content of H-FABP in a human body, belonging to the field of disease diagnosis and detection. Background technique [0002] Acute coronary syndrome (ACS) is one of the important acute events in coronary heart disease. It is divided into acute myocardial infarction (AMI), unstable angina (UA) and sudden cardiac death (SCD). It plays an important role in diagnosis and prognosis. Because ACS has the characteristics of acute onset and great harm, it is of great significance to select early markers with high specificity, early diagnosis and timely treatment for ACS patients. [0003] Heart-type fatty acid binding protein (H-FABP) is a new type of small molecular protein rich in the heart. It combines with two fatty acid molecules and participates in fat metabolism to provide energy for cardiomyocytes. It is highly specific for the heart. The concentrati...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/544G01N33/533
CPCG01N33/533G01N33/544G01N33/6887G01N2333/4712G01N2800/324
Inventor 徐林
Owner NANJING TZONE BIOLOGICAL SCI & TECH