Quick amplifying reagent kit and amplifying method of CYP2C19 gene

Abstract

The invention discloses a quick amplifying reagent kit and amplifying method of a CYP2C19 gene. The reagent kit comprises 3 pairs of amplifying primers, Taq enzymes, dNTPs and a buffered solution, wherein the amplifying primers are used for amplifying at least two SNP sites in a CYP2C19*2 type, a CYP2C19*3 type and a CYP2C19*17 type, and each pair of primers corresponds to upstream and upstream regions of one of SNP sites. A multiplex-PCR amplifying method is used, a plurality of important SNP sites of the CYP2C19 are amplified at the same time, the combination of sites to be amplified can beflexibly selected according to actual demands, and the effect that one reagent kit has many functions can be achieved; the primers in the reagent kit are high in specificity and low in mispairing rate, the length and the Tm value of the primers are reasonable to design, and amplified products are convenient to detect and separate. Multiple purpose products are amplified at a time, so that the amplifying efficiency can be greatly improved, and the amplifying cost can be reduced.

Description

technical field [0001] The invention relates to a rapid amplification kit and amplification method of CYP2C19 gene, belonging to the field of biology, in particular to the field of biological detection. Background technique [0002] Drug-metabolizing enzymes play an important role in the metabolism of drugs in vivo, and their activity directly determines the intensity and durability of drug effects. Cytochrome P450 (CYP450) is the main enzyme in the drug metabolism in the human body, and participates in the metabolism of 40% to 50% of marketed drugs, and its activity is an important factor determining the rate of drug metabolism. [0003] The CYP2C19 enzyme, called S-mephenytoin-4-hydroxylase, is one of the most important drug-metabolizing enzymes of the cytochrome P450 family (CYP450). Metabolism of many drugs such as , phenytoin and diazepam. The CYP2C19 enzyme is located in liver microsomes, and its coding gene is located at the q24.1-24.3 locus of human chromosome 10. ...

AI Technical Summary

Problems solved by technology

However, various detection methods are aimed at a single single nucleotide polymorphism site (SNP), and the detection time is long and the detection cost is high. However, the mutation frequency of the above three mutation sites is relatively high. Detection is not only time-consuming and labor-intensive, but also has low accuracy, making it difficult to give a qualitative conclusion on the site in a short time

Therefore, it is impossible to give the subjects accurate prediction of metabolic efficacy and medication guidance in real time and accurately.

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