ALDH2*2 genotype detecting kit and detecting method thereof

Abstract

The invention provides an ALDH2*2 genotype detecting kit and a detecting method thereof. The ALDH2*2 genotype detecting kit is characterized by comprising KOD DNA polymerase, an upstream primer, a downstream primer, a specific targeted fluorescent probe, a GG type positive reference, an AA type positive reference and an AG type positive reference. A GG homozygotic type DNA sequence is a section of linear recombinant DNA sequence which is designed by taking human ALDH2*2 gene 1510-position GG type as a template; an AA homozygotic type DNA sequence is a section of linear recombinant DNA sequence which is designed by taking human ALDH2*2 gene 1510-position AA type as a template; and the AG type positive reference is prepared by blending the GG type positive reference and the AA type positive reference in an isovolumetric manner. The detecting method can rapidly, simply and conveniently detect variation of the 1510 position of the ALDH2*2 gene.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to an ALDH2*2 genotype detection kit and a detection method thereof. The kit adopts PCR in vitro amplification method for qualitative detection of acetaldehyde dehydrogenase in human whole blood samples Genetic polymorphism at position 1510 of ALDH2*2. Background technique [0002] Mitochondrial aldehyde dehydrogenase 2 (ALDH2) has both acetaldehyde dehydrogenase and esterase activities, and participates in the metabolism of drugs such as ethanol and nitroglycerin. ALDH2 metabolically activates nitroglycerin to its active metabolite nitric oxide. ALDH2*2 (Glu504Lys, rs671) polymorphism leads to the substitution of glutamic acid at position 504 of the encoded protein by lysine, and the ALDH2 enzyme activity of individuals carrying the mutant allele (ALDH2*2) decreases, and the enzyme activity of heterozygous individuals is only 10% of wild-type individuals, mutant homozyg...

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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