Specific primer and liquid phase chip for SNP (Single Nucleotide Polymorphism) detection of CYP2C9 (Cytochrome P4502C9)

Abstract

The invention discloses a specific primer and liquid phase chip for SNP (Single Nucleotide Polymorphism) detection of CYP2C9 (Cytochrome P4502C9), wherein the liquid phase chip comprises an ASPE primer consisting of a tag sequence of 5' end and a specific primer of 3' end aiming at target gene mutation, a microsphere coated by the anti-tag sequence, and an amplification primer; the specific primer includes: SEQ ID NO.17 and SEQ ID NO.18 aiming at G98A SNP locus, SEQ ID NO.19 and SEQ ID NO.20 aiming at G173A SNP locus, SEQ ID NO.21 and SEQ ID NO.22 aiming at C90T SNP locus, SEQ ID NO.23 and SEQ ID NO.24 aiming at A162C SNP locus, SEQ ID NO.25 and SEQ ID NO.26 aiming at C72T SNP locus, SEQ ID NO.27 and SEQ ID NO.28 aiming at T102C SNP locus, SEQ ID NO.29 and SEQ ID NO.30 aiming at A77G SNP locus, and / or SEQ ID NO.31 and SEQ ID NO.32 aiming at A167C SNP locus. The matching rate between a detection result obtained by adopting the liquid phase chip for SNP detection of CYP2C9 and a sequencing method can be 100%.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a CYP2C9 gene SNP detection specific primer and a liquid phase chip. Background technique [0002] The main enzymes that metabolize drugs in the human body are the cytochrome P450 superfamily (Cytochrome P450 proteins, CYP). Metabolism of xenobiotics (including most clinical drugs). P450 enzymes can transfer electrons to oxidize heterologous substances through iron ions in non-covalently bonded heme, enhance the water solubility of heterologous substances, and make them easier to excrete. [0003] There are multiple subfamilies of CYP, among which CYP2C9 (Cytochrome P4502C9) is an important member of the second subfamily, accounting for 20% of the total amount of P450 proteins in liver microsomal. CYP2C9 can hydroxylate and metabolize many drugs with different properties, mainly acidic substrates. According to statistics, about 16% of cli...

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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