Detection method and detection kit for polymorphism of CYP2C9 and VKORC1 genes

Abstract

The invention relates to a detection method and detection kit for polymorphism of CYP2C9 and VKORC1 genes. The detection method comprises the following steps: (1) designing specific primers aiming atthe CYP2C9 and VKORC1 genes; (2) amplifying a target segment containing the CYP2C9 and VKORC1 genes by utilizing specific PCR (Polymerase Chain Reaction); (3) digesting a PCR product segment by utilizing restriction endonuclease; (4) carrying out single base extension reaction; (5) carrying out desalting and purification treatment; and (6) analyzing gene sequences of the target genes CYP2C9 and VKORC1 through a nucleic acid mass spectrometer. The detection method for the polymorphism of the CYP2C9 and VKORC1 genes has the advantages of high detection accuracy, high experiment repeatability, great flux and low cost. The invention further provides a detection kit for the polymorphism of the CYP2C9 and VKORC1 genes; and the detection kit comprises a specific primer pair for amplifying the CYP2C9 and VKORC1 genes. According to the detection kit for the polymorphism of the CYP2C9 and VKORC1 genes, an experiment flow is simplified, the operation time of workers is short, the difficulty is small and the automation degree of experiments is high.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a detection method and a detection kit for CYP2C9 and VKORC1 gene polymorphisms. Background technique [0002] Warfarin (warfarin) is a vitamin K antagonist oral anticoagulant widely used clinically at present, and is used for anticoagulation in venous thromboembolism, myocardial infarction, ischemic stroke, and atrial fibrillation after heart valve replacement Treatment and prevention. The clinical efficacy and adverse reactions of warfarin vary greatly among individuals, and the dose is difficult to control, especially in the initial stage of warfarin anticoagulant therapy, which can easily lead to serious bleeding complications. It is estimated that among patients taking warfarin, 15.2% of them experience bleeding side effects every year, of which fatal bleeding accounts for 3.5%, and the difference in the stable dose of warfarin among different individuals can r...

AI Technical Summary

Problems solved by technology

The most common method is the sequencing method, which has low cost and high throughput, but it takes a long time to operate and has low sensitivity, which is not suitable for rapid clinical detection; the high-resolution melting curve method has special requirements for equipment, and there are certain difficulties in clinical promotion.

However, the traditional fluorescent quantitative PCR method is widely used clinically, but this method generally uses the genotyping method to detect gene polymorphisms. This method has certain requirements for the number of samples and the distribution of polymorphisms. When the sample size is small, it cannot In addition, blood samples usually need to be extracted and purified to obtain nucleic acid as a template for fluorescent quantitative PCR amplification. Although the purified nucleic acid is of high purity, the cumbersome purification steps may not only cause cross-contamination between samples, At the same time, it is also easy to cause the loss of nucleic acid, thereby increasing the risk of subsequent fluorescent quantitative PCR failure

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