Detection kit and detection method for aspartate aminotransferase mitochondrial isozyme

An aspartate amino, mitochondrial isoenzyme technology, applied in the biological field, can solve the problems of low degree of automation of enzyme immunoassay, low m-AST content, unsuitable for large-scale operation, etc., and achieves low price and short time consumption. , easy to obtain effect

Pending Publication Date: 2019-08-02
天津中成佳益生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, there have been more and more studies on m-AST. The existing m-AST detection methods are mainly as follows: due to the low content of m-AST, the electrophoresis method requires a complicated dyeing process and a high-precision densitometer. Not suitable for clinical large-scale sample analysis; ion exchange column chromatography, this method is not suitable for large-scale operations; enzyme immunoassay has a low degree of automation; radioimmunoassay has radioactive contamination; Problems with routine project development
The above detection methods are cumbersome to operate, have low precision, poor quantitative ability, and take a long time, so they cannot be quickly detected.

Method used

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  • Detection kit and detection method for aspartate aminotransferase mitochondrial isozyme
  • Detection kit and detection method for aspartate aminotransferase mitochondrial isozyme
  • Detection kit and detection method for aspartate aminotransferase mitochondrial isozyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] A detection kit for aspartate aminotransferase mitochondrial isoenzyme, said kit is composed of independent reagent R1 and reagent R2;

[0052] The reagent R1 consists of the following components:

[0053] TRIS buffer: 12g / L;

[0054] Potassium L-aspartate: 50g / L;

[0055] Malate dehydrogenase: 3KU / L;

[0056] Anti-cellular AST monoclonal antibody: 20mg / L;

[0057] NADH: 7g / L;

[0058] The solvent is purified water;

[0059] Preparation of R1 reagent: Dissolve TRIS buffer in purified water and stir evenly to prepare R1 buffer. Add L-aspartate potassium, malate dehydrogenase, and anticellular AST monoclonal Antibody and NADH were sequentially dissolved in the prepared R1 buffer, mixed evenly and adjusted to pH 9.10 with HCL.

[0060] The reagent R2 consists of the following components:

[0061] TRIS buffer: 12g / L;

[0062]α-ketoglutaric acid: 11g / L;

[0063] Preparation of R2 reagent: Dissolve TRIS buffer in purified water, stir well to prepare R2 buffer, dissol...

Embodiment 2

[0073] A detection kit for aspartate aminotransferase mitochondrial isoenzyme, said kit is composed of independent reagent R1 and reagent R2;

[0074] The reagent R1 consists of the following components:

[0075] TRIS buffer: 12.114g / L;

[0076] Potassium L-aspartate: 60g / L;

[0077] Malate dehydrogenase: 2.5KU / L;

[0078] Anti-cellular AST monoclonal antibody: 25mg / L;

[0079] NADH: 8g / L;

[0080] The solvent is purified water;

[0081] Preparation of R1 reagent: Dissolve TRIS buffer in purified water and stir evenly to prepare R1 buffer. Add L-aspartate potassium, malate dehydrogenase, and anticellular AST monoclonal Antibody and NADH were dissolved in the prepared R1 buffer in turn, and after mixing evenly, the pH was adjusted to 9.20 with HCL.

[0082] The reagent R2 consists of the following components:

[0083] TRIS buffer: 12.5g / L;

[0084] α-ketoglutaric acid: 12g / L;

[0085] Preparation of R2 reagent: Dissolve TRIS buffer in purified water, stir well to prepa...

Embodiment 2

[0092] Table 2 embodiment 2 measurement parameters

[0093]

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PUM

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Abstract

The invention provides a detection kit and a detection method for an aspartate aminotransferase mitochondrial isozyme. The kit consists of a reagent R1 and a reagent R2 which are mutually independent;the reagent R1 is prepared from the following components: 12-13g/L of TRIS buffer, 40-60g/L of L-potassium aspartate, 2.5-4KU/L of malate dehydrogenase, 15mg/L-25mg/L of anti-cell type AST monoclonalantibody, and 6-8g/L of NADH; a solvent for the reagent R1 is purified water; the reagent R2 prepared from the following components: 12-13g/L of TRIS buffer and 10-12g/L of alpha-ketoglutaric acid; asolvent for the reagent R2 is purified water. The detection kit and the detection method provided by the invention have the advantages of good quantitative ability, short time consumption, rapid batch detection, good repeatability, good stability, simple operation and low price, and can be applied to multi-brand clinical biochemical analyzers with open channels.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a detection kit and a detection method for mitochondrial isozymes of aspartate aminotransferase. Background technique [0002] Aspartate aminotransferase (AST) is widely distributed in the heart, liver, lung, skeletal muscle, kidney, pancreas and other tissue cells. The highest content in cardiomyocytes, followed by liver cells. AST has two isozymes, one is the cytoplasmic isozyme of aspartate aminotransferase (s-AST) located in the cytoplasm; the other is the mitochondrial isozyme of aspartate aminotransferase located in the mitochondria of cells (m-AST). The changes in the concentrations of the two isoenzymes in serum have different clinical significance, among which the m-AST isoenzyme can objectively reflect the severity of some diseases and serve as an indicator for guiding treatment and prognosis of some diseases. In recent years, there have been more and more studies on m-A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/48
CPCC12Q1/48G01N2333/91188
Inventor 陈逸桐赵德明戴俊刘宏婧靳峰李园园吴凯杰
Owner 天津中成佳益生物科技有限公司
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