Single-nucleotide polymorphism (SNP) detection specific primer of CYP2C19 gene and liquid phase chip

Abstract

The invention discloses a single-nucleotide polymorphism (SNP) detection specific primer of the CYP2C19 gene and a liquid phase chip. The liquid phase chip contains an ASPE primer composed of a tag sequence on the 5'-end and specific primers on the 3'-end which aims at the target genetic mutation, microspheres covered by the anti-tag sequence and an amplification primer, wherein the specific primers contain SEQ ID NO.25 and SEQ ID NO.26, SEQ ID NO.27 and SEQ ID NO.28 aiming at the C194T SNP site, SEQ ID NO.29 and SEQ ID NO.30, SEQ ID NO.31 and SEQ ID NO.32, SEQ ID NO.33 and SEQ ID NO.34, SEQID NO.35 and SEQ ID NO.36, SEQ ID NO.37 and SEQ ID NO.38, SEQ ID NO.39 and SEQ ID NO.40, SEQ ID NO.41 and SEQ ID NO.42, SEQ ID NO.43 and SEQ IDNO.44, SEQ ID NO.45 and SEQ ID NO.46, and / or SEQ ID NO.47 and SEQ ID NO.48. The coincidence rate of the detection result of the SNP detection liquid phase chip of the CYP2C 19 gene and the detection result adopting a sequencing method is up to 100%.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a CYP2C19 gene SNP detection specific primer and a liquid phase chip. Background technique [0002] Cytochrome oxidase P4502C19 (CYP2C19), also known as S-mephenytoin hydroxylase, is an important liver microsomal enzyme. CYP2C19 exists in liver microsomes, and many endogenous substrates, environmental pollutants, and about 2% of clinical drugs are catalyzed by it. Studies have found that CYP2C19 gene polymorphisms can affect the metabolism of many important clinically used drugs, such as omeprazole, diazepam, imipramine, propranolol, etc., which is one of the reasons for the different metabolism of the same drug between individuals and races. [0003] CYP2C19 is located on chromosome 10q24, consists of 490 amino acids, and has a molecular weight of 55933kb. The entire sequence includes 9 exons and 5 introns. The sequence is clear, and its i...

AI Technical Summary

Problems solved by technology

Fluorescent quantitative PCR has the advantages of simple operation, quick results, and quantification. However, this technology has the disadvantages of easy sample contamination, cross-reaction, and high false positive rate; while PCR-RFLP method is based on restriction endonucleation caused by gene mutations. Changes in the enzyme recognition site, such as site loss or new site generation, a specific fragment is amplified by PCR, and then the amplified product is digested with a restriction endonuclease, and the size of the fragment is observed by electrophoresis. This method is used for Detection of gene mutations with altered restriction sites can directly determine the genotype, but this method cannot be used for the detection of gene mutations without new restriction sites

Again, the above methods all have limitations in detection throughput, and can only detect one mutation type at a time, which cannot meet the needs of practical applications.

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