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Kit and method for determining genotype of predetermined SNP (single nucleotide polymorphisms) locus of DNA (deoxyribonucleic acid) sample to be detected

A kit and sample technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems that need to be improved, and achieve the effects of high accuracy, convenient operation, and high sensitivity

Inactive Publication Date: 2015-10-14
湖北维达健基因技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, the detection and genotyping methods of the rs1057910 locus of the CYP2C9 gene still need to be improved at this stage

Method used

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  • Kit and method for determining genotype of predetermined SNP (single nucleotide polymorphisms) locus of DNA (deoxyribonucleic acid) sample to be detected
  • Kit and method for determining genotype of predetermined SNP (single nucleotide polymorphisms) locus of DNA (deoxyribonucleic acid) sample to be detected
  • Kit and method for determining genotype of predetermined SNP (single nucleotide polymorphisms) locus of DNA (deoxyribonucleic acid) sample to be detected

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Embodiment 1

[0051] According to the method for determining the genotype of the predetermined SNP site of the DNA sample to be tested according to the present invention, the genotype of the rs1057910 site of the CYP2C9 gene of 57 cases of the DNA sample to be tested (human peripheral blood DNA sample) is determined, and the specific steps are as follows:

[0052] 1. Design and synthesize the following primers and probes:

[0053] Targeting the rs1057910 site of the CYP2C9 gene:

[0054] F primer: 5'-CCTACACAGATGCTGTGG-3' (SEQ ID NO: 1);

[0055] R primer: 5'-GGGAATGAGATAGTTTCTGAA-3' (SEQ ID NO: 2);

[0056] FAM probe: FAM-AGGTCCAGAGATACATTGACCTTCTCC-MGB;

[0057] VIC probe: VIC-AGGTCCAGAGATACCTTGACCTTCTCC–MGB.

[0058] 2. Fluorescent quantitative PCR:

[0059] ABI Stepone fluorescent quantitative PCR instrument (Applied Biosystems, Applied Biotechnology Co., Ltd., USA) was used to perform fluorescent quantitative PCR on each DNA sample, and the detection of the SNP site of each DNA sam...

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Abstract

The invention discloses a kit and a method for determining the genotype of a predetermined SNP (single nucleotide polymorphisms) locus of a DNA (deoxyribonucleic acid) sample to be detected. The kit comprises a first nucleic acid molecule, a second nucleic acid molecule, a first probe and a second probe, wherein the 5' terminals of the probes are labelled with fluorescence reporter groups and the 3' terminals are labelled with fluorescence quenching groups; the fluorescence reporter groups of the first and second probes are different. The kit can be used for detection and genotyping of an rs1057910 locus of a CYP2C9 gene.

Description

technical field [0001] The invention relates to a kit and a method for determining the genotype of a predetermined SNP site of a DNA sample to be tested. Background technique [0002] Warfarin (Warfarin, also known as "warfarin") is the most commonly used coumarin-type oral strong anticoagulant drug in clinical practice, and is widely used in heart valve replacement, atrial fibrillation, pulmonary embolism, and stroke again (stroke) Prevention and treatment of deep arteriovenous thrombotic diseases. The chemical structure of the drug is 3-(a-phenylacetone)-4-hydroxycoumarin, which is a racemic mixture of two optical isomers R-type and S-type, wherein S isomer The anticoagulant effect of the isomer is 3 to 5 times stronger than that of the R isomer. The dose of warfarin varies greatly among different races and individuals, even 10 to 20 times. Factors affecting the dose of warfarin mainly include genetic factors, height, weight, age, sex, race, and drug interactions. Rese...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 徐谋胜曹旸袁亚燕王珊
Owner 湖北维达健基因技术有限公司
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