Reagent kit and method for using molecular beacon probe for detecting human CYP2C9 gene polymorphism and application thereof

Abstract

The invention provides a reagent kit for using a molecular beacon probe for detecting human CYP2C9 gene polymorphism. The reagent kit comprises the following nucleotide sequences, a primer pair and the probe for detection, wherein the primer pair is used for amplifying polymorphic sites of rs1799853 and rs1057910 of a CYP2C9 gene. The invention further discloses application on CYP2C9 gene polymorphic site detectionand amplification of the primer and the probe. The reagent kit for using the molecular beacon probe for detecting the human CYP2C9 gene polymorphism is easy to operate, precise in result and easy to interpret, polymorphism of the sites of rs1799853 and rs1057910 of the CYP2C9 gene can be detected rapidly in PCR. Compared with a sequencing method, a method for using the molecularbeacon probe for detecting the human CYP2C9 gene polymorphism does not need to conduct a series of follow-up treatment on PCR products, the PCR amplification and detection are carried out synchronously, and a cover does not need to be opened in the whole detection process, so that risks for contaminating the PCR products are reduced, the detection time is dramatically shortened, and the detectioncost is reduced.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a kit, a method and an application of a molecular beacon probe for detecting human CYP2C9 gene polymorphism. Background technique [0002] CYP2C9 is an important member of the second subfamily of cytochrome P450 enzymes (CYP), and is an important drug-metabolizing enzyme in the human body; CYP2C9 can hydroxylate and metabolize many drugs with different properties. According to statistics, about 16% of clinical drugs are composed of CYP2C9 Responsible for metabolism. CYP2C9 participates in the hydroxylation metabolism of various drugs such as anticoagulant drugs, anticonvulsant drugs, hypoglycemic drugs, non-steroidal antipyretic analgesic anti-inflammatory drugs, antihypertensive drugs and diuretics, among which warfarin, toluenesulfonate Both butylurea and phenytoin have narrow therapeutic indices. Changes in CYP2C9 activity can lead to large changes in the ...

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Problems solved by technology

[0004] At present, the methods for polymorphic site analysis mainly include fluorescent quantitative PCR (qPCR), PCR-restriction fragment length polymorphism (PCR-Restriction Length Polymorphism, PCR-RFLP), amplification block mutation system-PCR (Amplification refractory mutation system PCR, ARMS-PCR), etc., but these methods are not suitable for large-scale rapid detection of the population because of the many steps, time-consuming and laborious, or the need for PCR post-processing, which is prone to pollution

Sequencing is the gold standard for nucleic acid sequencing. Although accurate, the steps are cumbersome and costly

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