Kit for rapidly detecting warfarin individualized medication related gene SNP sites, and its detection method

Abstract

The invention relates to a kit for rapidly detecting warfarin individualized medication related gene SNP sites, and its detection method, and belongs to a genetic detection technology in the clinic detection technique of the biomedical field. The genetic typing of two polymorphic sites comprising CYP2C9*3(1061A / C) and VKORC1(-1639G / A) is rapidly and accurately carried out through human peripheral blood DNA drawing, specific PCR amplification by a Taq man probe and fluorescence signal analysis. The invention provides primers, probes and kits which are used for detecting the polymorphic sites, a use of the primers and the probes in the preparation of the kits, and a use of the determination of the warfarin application amount of a patient according to the detection results of the polymorphic sites. The determination use can effectively reduce the generation of the warfarin application amount related adverse events and prevent thrombotic diseases. The detection method can be used for the auxiliary diagnosis and treatment of various patients needing warfarin clinically.

Description

Technical field [0001] The invention relates to a method that can be used to quickly detect the SNP sites of genes related to warfarin individualized medication, specifically by extracting human peripheral blood DNA and performing fluorescence signal analysis after specific PCR amplification with Taq man probes. Fast and accurate genotyping of two polymorphic loci CYP2C9*3 (1061A / C) and VKORC1 (-1639G / A). The method can be used for auxiliary diagnosis and treatment of various patients who need to take warfarin clinically, and belongs to the gene detection technology in the clinical detection technology of the biomedical field. Background technique [0002] Warfarin is currently a widely used coumarin-based oral anticoagulant for the prevention and treatment of thromboembolic diseases such as prosthetic valve replacement, venous thrombosis (pulmonary embolism), atrial fibrillation and other oral anticoagulation treatments, and Reduce the risk of myocardial infarction recurrence a...

AI Technical Summary

Problems solved by technology

However, this method is complicated to operate, and when the sample size is large, it is easy to cause cross-contamination of PCR products, and it is prone to insufficient or excessive digestion, resulting in false negative or false positive results, and low reliability.

Although the specificity of fluorescent quantitative PCR and multiplex PCR methods has been improved, the principles of these two methods are still based on the principle of ordinary PCR. Factors such as the strength of fluorescent signals, primer specificity, and low-fidelity Taq enzymes can cause impact on results

The DNA sequencing method is relatively expensive and the operation process is cumbersome. In today's rapid human post-genome era, these methods cannot meet the current requirements for rapid and accurate analysis of VKORC1 (-1639G / A) and CYP2C9 (1075A / C) SNP loci. Require

Images