Quick genotyping method for guiding Warfarin utilization starting dose and kit

Abstract

The invention discloses a quick genotyping method for guiding Warfarin utilization starting dose and a kit and belongs to the gene detection technology in clinical detection technology in the biomedicine field. According to the quick genotyping method, the deoxyribonucleic acid (DNA) of a host cell gene group is extracted, and the gene types of CYP2C9*3(1061 A / C) and VKORCI (-1639 G / A) of a subject are measured. The CYP2C9 *3(1061 A / C) and VKORCI (-1639 G / A) genes have a nucleic acid sequence site shown as SEQ ID 1, wherein the site is one of the two most main sites associated with Warfarin metabolism. The invention provides the quick and accurate genotyping method and the associated kit. The method is quick and simple in operation, and low in cost, can be applied to prevention, auxiliary diagnosis and treatment on thrombotic associated diseases, and is conveniently popularized and used in basic medical institutions.

Description

technical field [0001] The present invention relates to a method capable of guiding the rapid genotyping of the initial dosage of warfarin. Specifically, by extracting human peripheral blood DNA, amplifying by PCR, and then performing high-resolution melting curve analysis, CYP2C9*3 (1061 A / C) and VKORC1 (-1639 G / A) polymorphic loci for rapid genotyping. 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 in the field of biomedicine. Background technique [0002] Warfarin is currently a widely used coumarin oral anticoagulant for the prevention and treatment of thromboembolic diseases such as oral anticoagulant therapy for artificial valve replacement, venous thrombosis (pulmonary embolism), atrial fibrillation, and Reduce the risk of recurrent myocardial infarction and thromboembolic death after myocardial infarction. However...

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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.

The operation process of the probe method is simple, the specificity of PCR amplification is high, but the cost of ordering probes is too high, and the probes are prone to failure after repeated freezing and thawing, and the feasibility of clinical application is low.

Although the DNA sequencing method is currently the gold standard for genetic diagnosis, the steps are cumbersome, the process is complicated, the reagents are expensive, and cross-contamination between samples is prone to occur, resulting in sequencing failures; in addition, the equipment of the sequencer is beyond that of a general clinical laboratory. Tolerance range

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