ARMS primer for detecting CYP2C9 gene polymorphism

Abstract

The invention discloses an ARMS primer for detecting CYP2C9 gene polymorphism. Aiming at sequence changes in every SNP of CYPC2C9*2(rs1799853, C430T, Arg144Cys) and CYP2C9*3(rs1057910, A1075C, Ile359Leu), the specificity ARMS primer and shared primers are designed. Mismatched base group (base group is marked with underline) is introduced to the second or third place of the 3' terminal end of the specificity ARMS primer so as to enlarge efficiency gap generating when specificity ARMS primer 3' terminal base are complementary and are not complementary to the SNP locus base on the DNA template, thus sharply improving specificity and accuracy of the detection.

Description

technical field [0001] The invention belongs to the field of life science and biotechnology, and in particular relates to ARMS primers for detecting mutations of exons 3 and 7 of CYP2C9 gene, which can be used for rapid detection of mutations of polymorphic sites of CYP2C9 gene by using ordinary PCR technology and gel electrophoresis. Background technique [0002] Human gene polymorphisms play an important role in elucidating the susceptibility and tolerance of the human body to diseases and poisons, the diversity of clinical manifestations of diseases, and the responsiveness to drug treatment. [0003] The CYP2C9 gene is located on human chromosome 10, with a total length of 83529bp and a total of 10 exons. CYP2C9 is an important member of the second subfamily of cytochrome P450 enzymes (CYP), accounting for 20% of the total amount of P450 proteins in liver microsomal. CYP2C9 participates in the hydroxylation metabolism of various drugs such as anticoagulant drugs, anticon...

AI Technical Summary

Problems solved by technology

Among them, DNA sequencing is the gold standard for genotyping, but this method has some limitations in clinical application: low detection sensitivity, time-consuming, complicated operation, easy contamination, low throughput, etc.

Restriction fragment length polymorphism is only applicable when there is an appropriate specific restriction endonuclease recognition sequence near the mutation site, and its application is limited

The hybridization chip detection method has the problems of high false positive rate, precise annealing temperature, and low specificity; the preparation of Taqman method probes and supporting equipment are expensive and the accuracy is not high

[0005] The amplification block mutation system uses the principle that Taq enzyme has no 3'→5' exonuclease activity, and when the 3' terminal base is mismatched, it cannot be effectively amplified

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