CYP2C19 and ABCB1 gene SNP detection liquid-phase chip and detection method thereof

Abstract

The invention discloses a CYP2C19 and ABCB1 gene SNP detection liquid-phase chip and a detection method thereof. The liquid-phase chip comprises microspheres and amplification primer, wherein the microspheres are coated with specific anti-tag sequences respectively; and the anti-tag sequences are selected from sequences from SEQ ID NO.7 to SEQ ID NO.12. As for three pairs of ASPE primers formed by specific sequences of SNP sites of a target gene and tag sequences of 5' ends, the specific sequences are SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 and SEQ ID NO.6 respectively. The fluorescent signal value detected by the liquid-phase chip is greatly improved, so that the detection sensitivity is further improved, the signal-to-noise ratio is strengthened, and the detection result is more accurate and reliable.

Description

technical field [0001] The invention belongs to the field of molecular biology and relates to medicine and biotechnology, in particular to a liquid phase chip for detecting CYP2C19 and ABCB1 gene SNP and a detection method thereof. Background technique [0002] Plavix (also known as Clopidogrel, Clopidogrel) is a new generation of platelet aggregation inhibitors, which was officially approved by the FDA in March 1998. Its combination therapy with aspirin is widely used in patients with acute coronary syndrome and percutaneous coronary intervention (PCI), and can effectively prevent myocardial infarction, stroke and vascular death. At present, tens of thousands of patients around the world are taking Plavix for a long time to prevent thrombosis after implanting coronary stents. [0003] Plavix is ​​a prodrug that needs to be metabolized by P450 enzymes to form active metabolites. Scientific research has proved that there are significant individual differences in the efficac...

AI Technical Summary

Problems solved by technology

At present, products capable of detecting CYP2C19 gene SNPs at home and abroad, such as CodeLink P450 of Amersham Bioscience (GE healthcare), AmpliChip P450 of Roche, etc., are mainly based on traditional solid-phase chips, which are expensive and have low sensitivity. poor repeatability

However, other PCR-based techniques for detecting SNPs, such as direct sequencing, semi-quantitative PCR, and PCR-single-strand conformational polymorphism (SSCP) detection, have low sensitivity, easy sample contamination, and high false positive rates. Due to the limitations of the detection throughput, ordinary PCR methods and fluorescent quantitative PCR cannot meet the clinical needs

However, the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis technology and the allelic difference analysis method based on TaqMan technology can only detect one mutation at a time, which is time-consuming and laborious.

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