Primer system for detecting gene SNP (single nucleotide polymorphism) related to warfarin dosage and application of primer system

Abstract

The invention discloses a primer system for detecting 7 gene polymorphic sites related to warfarin dosage. A product prepared based on the primer system can be used for realizing the detection of 7 gene polymorphic sites related to warfarin dosage. The product is used so as to detect the genetypes of a warfarin user at the 7 gene polymorphic sites; and the detection result is combined with other clinical indexes so that a reference can be provided for a clinical doctor to reasonably define the warfarin dosage, and the side effect of a medicament is avoided. According to the invention, the primer system can be used for simultaneously detecting the 7 gene polymorphic sites on different genes in one reaction system; and compared with technologies such as sequence measurement and real-time fluorescencent quantitative PCR (polymerase chain reaction), the primer system disclosed by the invention has the advantages of lower cost, simplicity and convenience in operation and improved accuracy and sensitivity.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a detection method and product for determining a gene polymorphic site (SNP) related to the dose of warfarin, specifically by using multiplex PCR technology, single base extension technology and mass spectrometry Technology, a method for detecting 7 gene polymorphic sites related to warfarin dosage and corresponding kits. Background technique [0002] Warfarin is currently the most widely used oral anticoagulant of dicoumarin derivatives. It is mainly used clinically for the prevention and treatment of thromboembolic diseases, and it also needs to be taken after surgical operations such as heart valve replacement and stent placement. Warfarin to reduce the incidence of venous thrombosis. However, the effective therapeutic range of warfarin is narrow and there are large differences in the dosage among different individuals. To achieve the same effect, the difference between high and lo...

AI Technical Summary

Problems solved by technology

Because the method is slow in operation, low in accuracy, easy to be contaminated clinically, and cannot perform multiple detections, it cannot solve the above technical problems

[0011] Chinese patent application 201010234597.8, "a kit for detecting SNP sites of genes related to warfarin individualized drug use and its PCR amplification method", involves PCR combined electrophoresis detection method, which can only detect 2 SNP sites , can not perform multiple detection, and its accuracy is low, and it is easy to be contaminated clinically, so it cannot solve the above technical problems

[0012] Chinese patent application 201110211124.0, "Kit for detecting SNP sites related to warfarin individualized drug use and its multiplex PCR amplification method and detection method", discloses a method for detecting 4 SNP sites by PCR combined with electrophoresis, Compared with the previous technology, this method has made some progress, but there are still a small number of detected SNP sites, and the inherent accuracy of electrophoresis display results is low, and the defects of easy clinical contamination, so it cannot solve the above technical problems

Since this method only involves two SNP sites, and fluorescent quantitative PCR detection has high cost of synthetic probes, process duplication and long time, although it is different from the above technologies, it cannot solve the above technical problems

[0014] Chinese patent application 201110328422.8, "a kit for detecting polymorphisms of VKORC1 and CYP2C9 genes" discloses a kit for detecting polymorphisms of warfarin-related genes, which involves DNA sequencing technology, although the detection area Each site in the sequence can be detected and may guide clinical medication, but the disadvantages of sequencing technology are low sensitivity, inability to perform multiple detections, and slow operation

[0015] In summary, the current technical problems are: the lack of methods and products that can simultaneously detect multiple genetic polymorphic sites related to the dose of warfarin at one time, common detection technologies, such as sequencing, real-time fluorescent quantitative PCR, etc., All sites need to be detected one by one. When there are many sites, the operation is complicated and the cost is high. Therefore, most of the existing detection programs are only for VKORC1 (rs9923231), CYP2C9*2 (rs1799853) and CYP2C9*3 ( rs1057910) and a few polymorphic sites

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