ADH2*2 gene type detection kit and detection method thereof

Abstract

The invention relates to an ADH2*2 gene type detection kit and a detection method thereof. The ADH2*2 gene type detection kit is characterized by comprising KOD DNA polymerase, an upstream primer, a downstream primer, a specific targeting fluorescence probe, a GG type positive reference, an AA type positive reference and an AG type positive reference, wherein a GG homozygotic type DNA sequence is a section of linear recombinant DNA sequence designed by taking a 143-site GG model of a human ADH2*2 gene as a template; an AA homozygotic type DNA sequence is a section of linear recombinant DNA sequence designed by taking a 143-site AA model of the human ADH2*2 gene as a template; and the AG type positive reference is prepared by mixing the GG type positive reference with the AA type positive reference in an isopyknic manner. By utilizing the detection method, the variation of the site 143 of the ADH2*2 gene can be rapidly, simply and conveniently detected.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to an ADH2*2 genotype detection kit and a detection method thereof. The kit adopts a PCR in vitro amplification method for the qualitative detection of acetaldehyde dehydrogenase in human whole blood samples Genetic polymorphism at position 143 of ADH2*2. Background technique [0002] The system of alcohol dehydrogenase (Alcohol dehydrogenase, referred to as ADH) is called ethanol: coenzyme I oxidoreductase (alcohol: NAD+ oxidoreductase), which exists in large quantities in human and animal livers, plant and microbial cells, and is a zinc-containing metal Enzymes with broad substrate specificity. In humans and mammals, alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) constitute an alcohol dehydrogenase system, which participates in the metabolism of alcohol in the body and is an important metabolic enzyme in humans and animals. Alcohol dehydrogenase ADH2...

AI Technical Summary

Problems solved by technology

Main disadvantages: low sensitivity, especially in the detection of somatic mutations in tumor tissue, when the target gene mutation ratio in the tissue is less than 20%, false negative results may occur; there are special requirements for reagents and instruments, and it is not easy to popularize; operation Complex, relatively high cost, slow speed, low throughput

Main disadvantages: There are special requirements for reagents and instruments, and it is not easy to popularize; the detection sensitivity is limited, and it is prone to false negatives for low-abundance somatic mutations (<3%) in tumor tissues; the sequencing length is only more than 10 bases, and cannot be detected Analysis of long fragments

When the molecular weight marker reaction tube has no bands or weak bands, the possible reasons include leaking wells, insufficient or invalid fluorescent dyes, too long electrophoresis time or too high voltage

The advantage of this method is that it is low in cost and can be carried out in ordinary laboratories; the disadvantage is that it is only suitable for qualitative determination of DNA insertion / deletion polymorphisms or fusion genes, and cannot be used for SNP detection

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