Primer probe composition and kit for detecting polymorphism of human CYP2C19 gene and application

Abstract

The invention discloses a primer probe composition and kit for detecting the polymorphism of a human CYP2C19 gene and application. The primer probe composition comprises three pairs of specific primers for amplifying CYP2C19*2, CYP2C19*3 and CYP2C19*17 sites and three specific fluorescent probes. The primers and the probes have high sensitivity, strong specificity and strong anti-interference capability; the polymorphism of the gene is detected by adopting a manner combining asymmetric PCR (Polymerase Chain Reaction) amplification and a fluorescent probe melting curve analysis technology; different gene types can be effectively distinguished according to the quantity of melting peaks and Tm value and result interpretation is convenient, clear and objective. Single-tube sampling can be usedfor detecting 6 mutation types of 3 gene sites at the same time; the operation is simple and convenient and the detection efficiency is improved; a large batch of samples can be detected and clinicaloperation is facilitated.

Description

technical field [0001] The present invention relates to a primer, probe, kit and application for detecting human CYP2C19 gene polymorphism, in particular to a primer for detecting human CYP2C19 gene *2, *3, *17 polymorphism The probe composition, kit and application belong to the technical field of molecular biology. Background technique [0002] Clopidogrel (trade name: Plavix) is a novel thienopyridine derivative that irreversibly inhibits platelet aggregation by selectively binding to the ADP receptor coupled to adenylate cyclase on the surface of platelets. Clopidogrel is currently the most commonly used antiplatelet drug in the world because it can effectively reduce the risk and mortality of diseases such as myocardial infarction, ischemic stroke and acute coronary syndrome, and has good safety. drug. However, 4% to 40% of patients will develop resistance to clopidogrel, leading to adverse cardiovascular events, and the incidence of adverse clinical events in Asian p...

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Problems solved by technology

Among them, the most commonly used sequencing method can directly detect the location and type of the mutation site, but the method has cumbersome operation steps, a long detection cycle, and the amplification product is prone to contamination; the chip method involves gene-specific amplification, hybridization, detection, etc. High-throughput analysis can be carried out in one step, but the cost is high, the detection steps are complicated, and there are certain requirements for the number of samples; the high-resolution melting curve method has simple steps and does not require post-amplification processing, but it does not contain specificity. Allele specific amplification method uses ARMS primers for specific amplification, the operation method is simple and no post-amplification treatment is required, but the primer design is difficult Optimization, strict detection conditions, prone to false positives due to primer mismatches in actual operation

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