CYP3A4 gene SNP detection specific primer, liquid-phase chip and method

Abstract

The invention discloses a CYP3A4 gene SNP detection specific primer, a liquid-phase chip and a method; and the liquid-phase chip comprises wild-type and mutant ASPE primers which are respectively designed for each type of mutation points, microspheres which are respectively coated with specific anti-tag sequences, and the primer which is used for amplifying a CYP3A4 gene target sequence with CYP3A4*1B, CYP3A4*2, CYP3A4*3, CYP3A4*4, CYP3A4*5, CYP3A4*6, CYP3A4*15, CYP3A4*17, CYP3A4*18 and / or CYP3A4*19SNP sites. The CYP3A4 gene SNP detection liquid-phase chip has very good signal-noise ratio, and a designed probe and the anti-tag sequences essentially have no cross reaction. The designed ASPE primers have very good specificity and can accurately distinguish all types of mutation points. The detection method has simple steps and ten types of SNP sites can be detected at one step, so that the operation is convenient, thereby preventing a plurality of uncertain factors in a plurality of operation processes, and greatly improving the detection accuracy rate.

Description

technical field The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a CYP3A4 gene SNP detection specific primer, a liquid phase chip and a method. Background technique Cytochrome P450 (cytochrome P450, cytochrome CYPs, P450 for short) is a class of proteases of the B family cytochrome superfamily with heme as the prosthetic group, and exists on the endoplasmic reticulum membrane of cells. P450 enzymes are the most important group of metabolic enzymes, involved in the metabolism of carcinogens and clinical therapeutic drugs. In the CYP450 superfamily, CYP1A2, CYP2A6, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 are related to the metabolism and drug-drug interactions of more than 95% of the drugs currently on the market. CYP3A isoenzymes are involved in the metabolism of tobacco carcinogens and steroids, including CYP3A4, CYP3A5, CYP3A7 and CYP3A43, and there are significant inter-individual differences in the transcription...

AI Technical Summary

Problems solved by technology

At present, there are a few reports on methods for detecting CYP3A4 gene polymorphism at home and abroad, mainly based on traditional solid-phase chips and PCR. Solid-phase chips are expensive, and the sensitivity is not high, and the reproducibility of test results is poor.

However, other PCR-based SNPs detection technologies, such as direct sequencing, semi-quantitative PCR technology, PCR-single-strand conformation polymorphism (SSCP) detection, etc., have the disadvantages of low sensitivity, easy sample contamination, and high false positive rate.

Ordinary PCR methods and fluorescent quantitative PCR cannot meet clinical needs due to the limitation of detection throughput, while polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis technology and allelic difference analysis based on TaqMan technology The method can only detect one mutation at a time, which is time-consuming and laborious

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