Primer and probe composition for detecting polymorphism of human CYP2C9 and VKORC1 genes, kit and application

Abstract

The invention discloses a primer and probe composition for detecting polymorphism of human CYP2C9 and VKORC1 genes, a kit and application. The primer and probe composition comprises three pairs of specific primers for amplifying CYP2C9*2, CYP2C9*3 and VKORC1 sites, and three specific fluorescent probes. The primers and the probes, disclosed by the invention, have high sensitivity, strong specificity and strong anti-interference capability; a manner combining an asymmetric PCR (Polymerase Chain Reaction) amplification and fluorescent probe melting curve analysis technologies is used for detecting the gene polymorphism; different gene types can be effectively distinguished according to the quantity and Tm value of a melting peak; a result is convenient, clear and objective to judge and read.Single-tube sampling can be used for simultaneously detecting 6 mutation types of 3 gene sites; the primer and probe composition is simple and convenient to operate and the detection efficiency is improved; a large batch of samples can be detected and clinical operation is facilitated.

Description

technical field [0001] The invention relates to a primer probe composition, kit and application for detecting human CYP2C9 and VKORC1 gene polymorphisms, belonging to the technical field of molecular biology. Background technique [0002] Warfarin is an indirect-acting coumarin-based oral anticoagulant. It inhibits the synthesis of coagulation factors II, VII, IX, and X by inhibiting vitamin K in liver cells, thereby exerting an anticoagulant effect. It is the most widely used oral anticoagulant in clinical practice. Drugs, which can prevent the formation and development of thrombus, are suitable for the prevention and treatment of thromboembolic diseases, such as the treatment of thromboembolic phlebitis, reducing the morbidity and mortality of pulmonary embolism, reducing major surgical operations, rheumatic heart disease, and hip fixation The incidence of venous thrombosis in surgery, artificial heart valve replacement surgery, etc. And can be used as an adjuvant drug fo...

AI Technical Summary

Problems solved by technology

Among them, the most commonly used sequencing method can directly detect the position 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 Difficult to optimize, strict detection conditions, prone to false positives due to primer mismatch in actual operation

Images