Quick genotyping method for guiding Warfarin utilization starting dose and kit

A technology of genotyping and kits, applied in biochemical equipment and methods, microbiological determination/inspection, etc., can solve the problems of low reliability, complicated operation, insufficient, etc., and achieve low cost, high sensitivity and specificity , the effect of easy operation

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A technology of genotyping and kits, applied in biochemical equipment and methods, microbiological determination/inspection, etc., can solve the problems of low reliability, complicated operation, insufficient, etc., and achieve low cost, high sensitivity and specificity , the effect of easy operation

CN102605076AActive Publication Date: 2012-07-25珠海赛乐奇医学检验有限公司

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Embodiment 1

[0019] Example 1 Detection of a kit that can guide warfarin to use the initial dose of rapid genotyping

[0020] 2. Kit components

[0021] 1. PCR amplification reagents

[0022]

[0023] F Primer 5'-ACGTGTGATTGGCAGAAACCG-3' (SEQ ID 2)

[0024] R primer 5'-GGCAGGCTGGTGGGGAGA-3' (SEQ ID 3)

[0025] F primer 5'-CAAAATGCTAGGATTATAGGCGT-3' (SEQ ID 4)

[0026] R primer 5'-CTGGGAAGTCAAGCAAGAGAAG-3' (SEQ ID 5)

[0027] 2. PCR reaction parameters:

[0028]

[0029] PCR instrument: Rotor-Gene 6000 TM (Australia Corbett Research Company), using Rotor-Gene Q Series Software 1.7 software for analysis.

[0030] 3. Test results

[0031] After the PCR reaction, the product was directly subjected to melting curve reaction and analyzed with Rotor-Gene Q Series Software 1.7 software. The results are shown in the figure: a is the melting curve, and b is based on VKORC1 (-1639 G / A) The conversion diagram of the melting curve; the diagram c is the melting curve diagram, and the diagr...

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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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Application Information

Patent Timeline

25 Jul 2012

Publication

CN102605076A

IPC: C12Q1/68

Inventors: 汪道文; 崔广林