Primers and probes for detecting ALDH2, and detection method

Abstract

The invention provides primers and probes for detecting an ALDH2 (aldehyde dehydrogenase gene), and a detection method for detecting the ALDH2. The detection method is characterized in that primers and probes of heterozygous ALDH2*1 / *2, homozygous ALDH2*2 / *2 and internal reference gene beta-globin are used for preparing PCR (Polymerase Chain Reaction) liquid; human whole blood sample DNA (Deoxyribonucleic Acid) is added; and the polymorphism of the ALDH2 is detected through a fluorescent PCR instrument. According to the method, the specificity detection probe is used for recognizing a target gene, and the accuracy is high. Meanwhile, a target sequence is controlled in a duplex way by the primers and the probes; the specificity is good; the false positive rate is low; the sensitivity is 0.01 percent, i.e., the detection can be realized when one cell with the polymorphism of the ALDH2*1 / *2 of the ALDH2 and the ALDH2*2 / *2 of the ALDH2 exists in 10,000 cells; the amplification and the detection can be performed in the same tube; the cover opening is not needed; the pollution cannot easily occur; meanwhile, the amplification and the detection can be completed in one step; later-stage processing is not needed; and the whole-process monitoring is realized. A kit provided by the embodiment of the invention introduces an internal positive control quality control system; the whole-process quality monitoring is realized in the detection process; the false positive result or the false negative result is effectively avoided; the speed is high; and the process can be completed in 100 minutes.

Description

Technical field: [0001] The invention relates to primers, probes and a detection method for detecting ALDH2 gene polymorphism in human blood samples, belonging to the field of molecular biology. Background technique: [0002] Aldehyde dehydrogenase (aldehydedehydrogenasegene, ALDH) is a group of NAD (P) + dependent enzymes, widely involved in the metabolic process of ethanol, amino acids, biogenic amines, vitamins, cholesterol and lipids produced by the aldehydes The oxidation reaction becomes the corresponding carboxylic acid, which is of great significance to reduce the toxic effect of aldehydes on the body. There are 19 isoenzymes of ALDH, the common ones are ALDH1~ALDH4, among which ALDH2 located in the mitochondria has the strongest activity and plays a major role in removing aldehydes in the body. The change of ALDH2 activity is related to the polymorphism of ALDH2 gene, and the mutant allele forms three genotypes in the population: wild homozygous ALDH2*1 / *1 with nor...

AI Technical Summary

Problems solved by technology

[0007] At present, the ALDH2 gene detection methods commonly used in the market include gene chip method and PCR-RFLP. Expensive, complex operation, low detection sensitivity and poor repeatability, etc.

PCR-restriction fragment length polymorphism (PCR-RFLP) has not been widely used due to its disadvantages such as cumbersome operation, long time consumption, and low reliability of results.

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