Kit and detection method thereof for SNP detection of aspirin individualized medication related genes

A technology of aspirin and detection method, which is applied in the field of aspirin individualized drug-related gene SNP detection, can solve the problems of high technical operation requirements, low detection throughput, low detection sensitivity, etc., and achieve the effect of solving expensive prices

Inactive Publication Date: 2017-02-22
中源协和基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Sequencing technology is recognized as the gold standard for detecting gene mutations, but it is difficult to form due to high equipment costs, long detection cycle, low detection throughput, low detection sensitivity, high technical requirements for experimenters, and cumbersome results determination steps. Commercial diagnostic products; the detection sensitivity of restriction fragment length polymorphism analysis is also not high, and the operation steps are cumbersome. The detection results still need to be verified again by the next generation sequencing method, especially when the sample size is large, it is easy to cause crossover of PCR products Contamination and prone to insufficient or excessive enzyme digesti

Method used

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  • Kit and detection method thereof for SNP detection of aspirin individualized medication related genes
  • Kit and detection method thereof for SNP detection of aspirin individualized medication related genes
  • Kit and detection method thereof for SNP detection of aspirin individualized medication related genes

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Effect test

Embodiment 1

[0070] A kit for instructing SNP detection of aspirin individualized medication-related genes comprises: including 2X qPCR mixed reaction solution, primer mix for detecting rs10306114 site of PTGS1 gene, primer mix for detecting rs1057910 site of CYP2C9 gene, and primer mix for detecting rs28360521 site of CYP2D6 gene The primer mix for the dot and the primer mix for detecting the rs12248560 site of the CYP2C19 gene.

[0071] The sequences of two specific forward primers and one reverse primer for detecting the rs10306114 site of the PTGS1 gene are as follows:

[0072] A allele forward primer: GAAGGTGACCAAGTTCATGCTGAGCACCTACTACATGCTGGA SEQ ID NO.3

[0073] G allele forward primer: GAAGGTCGGAGTCAACGGATTGAGCACCTACTACATGCTGGG SEQ ID NO.4

[0074] Reverse primer: TGCACACAAATCTCCTGGTGCAGT SEQ ID NO.5

[0075] The sequences of two specific forward primers and one reverse primer for detecting the rs1057910 site of the CYP2C9 gene are as follows:

[0076] A allele forward primer: G...

Embodiment 2

[0089] A PCR amplification method adopted by a kit for instructing the detection of SNP related genes of individualized medicine of aspirin comprises the following steps:

[0090] Step 1: Extraction of genomic DNA from the sample to be tested

[0091] (1) A blood sample is drawn from the patient.

[0092] (2) DNA was obtained from blood, and the TGuide blood genomic DNA extraction kit (OSR-M102) produced by Tiangen Biochemical (Beijing) Co., Ltd. was used for extraction on its matching TGuide M16 nucleic acid automatic extraction instrument.

[0093] The extraction process is as follows:

[0094] a. Add 200μl / 400μl mammalian whole blood sample to the sample tube, and add 10μl / 20μl proteinase K to mix well.

[0095] b. Place the sample tube in hole 4 of the T-frame. Run program number 102 (whole blood genomic DNA extraction program) and select the corresponding sample volume and final elution volume.

[0096] Step 2: Amplify the DNA

[0097] A 10ng / 10ul reaction system was...

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Abstract

The invention discloses a kit and a detection method thereof for SNP detection of aspirin individualized medication related genes. The kit consists of two forward primers and a reverse primer for detecting rs10306114 site of a PTGS1 gene, two forward primers and a reverse primer for detecting rs1057910 site of a CYP2C9 gene, two forward primers and a reverse primer for detecting rs2836051 site of a CYP2D6 gene, and two forward primers and a reverse primer for detecting rs12248560 site of a CYP2C19 gene. With the application of the kit provided by the invention, high-throughput detection can be conducted on the four sites and a genotyping results can be intuitively distinguished through software, so that the purposes of conducting quantitative control on aspirin measurement and reducing side effects to the greatest extent are achieved.

Description

technical field [0001] The invention relates to a kit for multiplex gene detection and a detection method thereof, in particular to a kit for detection of SNP of a gene related to individualized medicine of aspirin and a detection method thereof. Background technique [0002] Aspirin is the basic drug for the treatment of acute coronary syndrome and antithrombotic after percutaneous coronary intervention, and is widely used in the primary and secondary prevention of cardiovascular and cerebrovascular diseases. It is clinically found that some patients cannot effectively inhibit platelet activity despite long-term low-dose aspirin, that is, aspirin resistance. [0003] PTGS1 encodes COX-1, and PTGS1 gene polymorphisms can cause base substitutions and changes in promoter junctions, significantly affect the function of introns or exons, change the conformation of COX-1 protein, and make aspirin effective against COX-1. The sensitivity of aspirin is not uniform, thereby affecti...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q1/6858C12Q2600/106C12Q2600/156C12Q2600/16C12Q2531/113C12Q2537/143C12Q2527/107
Inventor 李同恩刘志霈王建刚孙颖朱兵王秉孝陶然王晶晶
Owner 中源协和基因科技有限公司
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