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Primer and probe for detecting ACE gene polymorphisms, application, kit and detection method

A gene polymorphism and detection kit technology, applied in the fields of molecular biology and medicine, can solve problems such as less attention to the long-term effects of gene mutations, improve accuracy and detection efficiency, reduce operational errors, and enhance specificity Effect

Pending Publication Date: 2018-10-23
HANGZHOU KMB BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the research on hypertension based on the pharmacogenomics group mainly focuses on the relationship between gene diversity and drug antihypertensive efficacy, and the influence of genetic differences caused by different races, ages, genders, diets, and living environments on the antihypertensive efficacy of drugs, etc. Less attention has been paid to the long-term effects of gene mutations on antihypertensive drugs

Method used

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  • Primer and probe for detecting ACE gene polymorphisms, application, kit and detection method
  • Primer and probe for detecting ACE gene polymorphisms, application, kit and detection method
  • Primer and probe for detecting ACE gene polymorphisms, application, kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1 Utilizes the detection kit of fluorescent quantitative PCR detection system

[0061] The detection kit includes amplification primers, probes, and other supporting reagents for fluorescent quantitative PCR detection.

[0062] Amplification primers and probes are for seven polymorphisms of CYP2C9*3(rs1057910), CYP2D6(rs1065852), CYP3A5*3(rs776746), ADRB1(rs1801253), AGTR1(rs5186), ACE(rs4646994) and NPPA(rs5065) Sex site design.

[0063] 1. Design and synthesis of amplification primers

[0064] Based on the region of rs1057910 in the CYP2C9*3 gene, the region of rs1065852 in the CYP2D6 gene, the region of rs776746 in the CYP3A5*3 gene, the region of rs1801253 in the ADRB1 gene, and the rs5186 in the AGTR1 gene The region of the site, the region of the rs4646994 site in the ACE gene, and the region of the rs5065 site in the NPPA gene, design amplification primers, wherein,

[0065] Primer sequences for amplifying CYP2C9*3 (rs1057910):

[0066] CYP2C9*3 (r...

Embodiment 2

[0120] Embodiment 2 fluorescent quantitative PCR detection method

[0121] The fluorescent quantitative PCR detection method for detecting the 7 polymorphic sites comprises the following steps:

[0122] (a) extracting the genomic DNA in the sample;

[0123] (b) Amplify and detect the gene polymorphism of the region containing the rs1057910 site in the CYP2C9*3 gene, the gene polymorphism of the region containing the rs1065852 site in the CYP2D6 gene, and the region containing the rs776746 site in the CYP3A5*3 gene Gene polymorphism in the region of ADRB1 gene including rs1801253 site, gene polymorphism in the region of AGTR1 gene including rs5186 site, gene polymorphism in the region of ACE gene including rs4646994 site 1. Gene polymorphism in the region including the rs5065 site in the NPPA gene.

[0124] The extraction methods of genomic DNA include: traditional phenol chloroform extraction, commercial silica gel membrane extraction kit, and commercial magnetic bead method...

Embodiment 3

[0130] Embodiment 3 adopts fluorescent quantitative PCR method to analyze clinical samples

[0131] Collect 150 cases of clinical saliva samples and 150 cases of clinical blood samples, utilize the commercialized magnetic bead method extraction kit to carry out the extraction of genomic DNA, then utilize the method for embodiment 1 and embodiment 2 to carry out the genotype detection of 7 genes. At the same time, the extracted DNA was genotyped using the first-generation sequencing method, and the genotyping results of this method were compared with the genotyping results of the first-generation sequencing. Among them, the comparative test results of CYP2C9*3(rs1057910) are shown in Table 3; the comparative test results of CYP2D6(rs1065852) are shown in Table 4; the comparative test results of CYP3A5*3(rs776746) are shown in Table 5; the comparative test results of ADRB1(rs1801253) As shown in Table 6; the comparative test results of AGTR1 (rs5186) are shown in Table 7; the co...

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Abstract

The present invention relates to genetic testing for guiding the guidance of hypertension medication, and discloses a primer and a probe for detecting ACE (rs4646994) polymorphisms and application thereof. The primer and probe can effectively detect insertion / deletion mutant ACE (rs4646994) genes on a fluorescent quantitation PCR platform, the operation method is simple and easy, that is, the riskof pollution is reduced, the efficiency is improved, a new simple judgment way is provided for clinical hypertension medication, and the primer and the probe have very high clinical value.

Description

technical field [0001] The invention relates to the fields of molecular biology and medicine, in particular to a detection composition, a kit and a detection method for guiding the SNP site associated with hypertension medication. Background technique [0002] At present, there are 5 categories of antihypertensive drugs commonly used in clinical practice: diuretics, angiotensin converting enzyme inhibitors, angiotensin receptor blockers, β-receptor blockers, and calcium ion antagonists. Drug metabolism in the body involves a variety of drug-metabolizing enzymes, transporters, and receptors, and their genetic polymorphisms ultimately lead to the therapeutic effect, adverse development, and risk factors when patients take the same drug in the same way. There are obvious individual differences in drug tolerance and other aspects. [0003] At present, the research on hypertension based on the pharmacogenomics group mainly focuses on the relationship between gene diversity...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6883C12Q2600/106C12Q2600/136C12Q2600/156
Inventor 林源吉丁佳女
Owner HANGZHOU KMB BIOTECH
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