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Genetic typing detection primer group related to cardiovascular and cerebrovascular disease drugs, kit and detection method

A cardiovascular and cerebrovascular disease, detection kit technology, applied in the fields of genetic engineering and molecular biology, can solve the problems of low detection sensitivity, high false positive rate, small throughput, etc., achieve high throughput, reduce complexity, and detect short cycle effect

Pending Publication Date: 2019-03-01
WUHAN CONSIDERIN GENE & HEALTH TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0016] The purpose of the present invention is to provide a primer set, kit and detection method for detecting drug-related genotyping of cardiovascular and cerebrovascular diseases, aiming at solving the problem of low sensitivity and high false positive rate of drug-related genotyping for cardiovascular and cerebrovascular diseases in the prior art , long cycle, low flux technical problems

Method used

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  • Genetic typing detection primer group related to cardiovascular and cerebrovascular disease drugs, kit and detection method
  • Genetic typing detection primer group related to cardiovascular and cerebrovascular disease drugs, kit and detection method
  • Genetic typing detection primer group related to cardiovascular and cerebrovascular disease drugs, kit and detection method

Examples

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Effect test

no. 1 example

[0064]The present invention proposes the first embodiment, a primer set for detection of cardiovascular and cerebrovascular disease drug-related genotyping, which is used to specifically amplify nucleic acid fragments containing sites to be tested, and the multiple cardiovascular and cerebrovascular disease drug-related The gene sites to be tested include CYP2C9 gene rs1799853 site, CYP2C9 gene rs1057910 site, CYP4F2 gene rs2108622 site, VKORC1 gene rs9923231 site, CYP2C19 gene rs4244285 site, CYP2C19 gene rs4986893 site, CYP2D6 gene rs2 site 06 rs671 site, MTHFR gene rs1801133 site, SLCO1B1 gene rs4149056 site, SLCO1B1 gene rs2306283 site, ABCG2 gene rs2231142 site, the primer set includes rs1799853 primer set, rs1057910 primer set, rs2108622 primer set, rs99232431 primer set 28rs At least one of the primer set, rs4986893 primer set, rs1065852 primer set, rs671 primer set, rs1801133 primer set, rs4149056 primer set, rs2306283 primer set, and rs2231142 primer set, wherein

[0...

no. 3 example

[0091] The present invention proposes a third embodiment, a drug-related genotyping detection method for cardiovascular and cerebrovascular diseases, comprising the following steps:

[0092] A. Using the primer set, the CYP2C9 gene rs1799853 locus, CYP2C9 gene rs1057910 locus, CYP4F2 gene rs2108622 locus, VKORC1 gene rs9923231 locus, CYP2C19 gene rs4244285 locus, CYP2C19 gene rs4986893 locus, CYP06585rs , ALDH2 gene rs671 locus, MTHFR gene rs1801133 locus, SLCO1B1 gene rs4149056 locus, SLCO1B1 gene rs2306283 locus, and ABCG2 gene rs2231142 locus are amplified to obtain amplified products; the primer set Including rs1799853 primer set, rs1057910 primer set, rs2108622 primer set, rs9923231 primer set, rs4244285 primer set, rs4986893 primer set, rs1065852 primer set, rs671 primer set, rs1801133 primer set, rs23064283 in primer set 1 and rs23064283 at least one,

[0093] The rs1799853 primer set includes rs1799853 upstream primer SEQ ID NO:1 and rs1799853 downstream primer SEQ ID...

no. 4 example

[0122] The present invention provides a fourth embodiment, a method for detecting genotypes related to drugs for cardiovascular and cerebrovascular diseases, using the blood genome DNA of two people as a template to detect the rs1799853 site of the CYP2C9 gene, the rs1057910 site of the CYP2C9 gene, and the CYP4F2 Gene rs2108622, VKORC1 gene rs9923231, CYP2C19 gene rs4244285, CYP2C19 gene rs4986893, CYP2D6 gene rs1065852, ALDH2 gene rs671, MTHFR gene rs1801133, SLCO2B1 gene rs414rs2CO5 locus, ABCG2 gene rs2231142 locus. It specifically includes steps A-G.

[0123] Step A. For the above two samples, two reaction systems were prepared, and the primer sets of the first embodiment were used to respectively target the rs1799853 site of the CYP2C9 gene, the rs1057910 site of the CYP2C9 gene, the rs2108622 site of the CYP4F2 gene, and the rs9923231 site of the VKORC1 gene of different samples. point, CYP2C19 gene rs4244285 site, CYP2C19 gene rs4986893 site, CYP2D6 gene rs1065852 sit...

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Abstract

The invention relates to the field of genetic engineering and molecular biology, and provides a genetic typing detection primer group related to cardiovascular and cerebrovascular disease drugs, whichis used for specific amplification of nucleic acid fragments containing a site to be measured, the site to be measured comprises an rs1799853 site of a CYP2C9 gene, an rs1057910 site of the CYP2C9 gene, an rs2108622 site of a CYP4F2 gene and the like, the primer group comprises an rs1799853 primer group, an rs1057910 primer group, and an rs2108622 primer group and the like. The invention also provides a gene detection kit and a detection method. The gene detection primer group can carry out specific amplification on the nucleic acid fragments containing the sites, so that the sensitivity of gene mutation detection is high and the false positive rate is low. According to the invention, a second generation high-throughput gene sequencing technology is used to detect the genotypes of the sites to be measured, which makes the detection cycle short and the flux high.

Description

technical field [0001] The invention relates to the fields of genetic engineering and molecular biology, and more specifically relates to a drug-related genotyping detection primer set, kit and detection method for cardiovascular and cerebrovascular diseases. Background technique [0002] Studies have shown that currently widely used drugs for cardiovascular and cerebrovascular diseases, such as warfarin, losartan, clopidogrel, prasugrel, ticagrelor, metoprolol, folic acid, etc., have different treatments for different individual genotypes Effect. [0003] CYP4F2*3 polymorphisms are associated with warfarin steady-state dose. CYP4F2*3 polymorphism can lead to decreased enzyme activity, increased vitamin K concentration, and enhanced anticoagulant effect of warfarin. Individuals carrying the CYP4F2*3 allele have a significantly increased risk of bleeding on warfarin. The CPIC guidelines recommend reducing the dosage of warfarin and coumarin anticoagulants (acenocoumarol, p...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6883C12Q2600/156C12Q2535/122C12Q2525/191C12Q2563/143C12Q2563/149
Inventor 盛司潼廖玉声张晓英郑乔骏
Owner WUHAN CONSIDERIN GENE & HEALTH TECH CO LTD
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