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Detection composition for guiding hypertension medication, applications of detection composition, kit and detection method

A detection kit and technology for hypertension, applied in the fields of molecular biology and medicine, can solve problems such as less attention to the long-term effects of gene mutations, and achieve the effects of improving accuracy and detection efficiency, reducing detection costs, and enhancing specificity

Active Publication Date: 2017-02-01
HANGZHOU KMB BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, 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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  • Detection composition for guiding hypertension medication, applications of detection composition, kit and detection method
  • Detection composition for guiding hypertension medication, applications of detection composition, kit and detection method
  • Detection composition for guiding hypertension medication, applications of detection composition, kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Detection kit using fluorescent quantitative PCR detection system

[0064] The detection kit includes amplification primers, probes, and other supporting reagents for fluorescent quantitative PCR detection.

[0065] 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.

[0066] 1. Design and synthesis of amplification primers

[0067] 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,

[0068] Primer sequences for amplifying CYP2C9*3 (rs1057910):

[0069] CYP2C9*3 (rs1057910) ups...

Embodiment 2

[0123] Example 2 Fluorescent quantitative PCR detection method

[0124] The fluorescent quantitative PCR detection method for detecting the 7 polymorphic sites comprises the following steps:

[0125] (a) extracting the genomic DNA in the sample;

[0126] (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 including rs1801253 in the ADRB1 gene, gene polymorphism in the region including rs5186 in the AGTR1 gene, gene polymorphism in the region including rs4646994 in the ACE gene 1. Gene polymorphism in the region including the rs5065 site in the NPPA gene.

[0127] Genomic DNA extraction methods include: traditional phenol-chloroform extraction, commercial silica gel membrane extraction kits, and commercial magnetic bead extraction kits. ...

Embodiment 3

[0133] Example 3 Analysis of Clinical Samples Using Fluorescent Quantitative PCR

[0134] Collect 150 clinical saliva samples and 150 clinical blood samples, use a commercial magnetic bead method extraction kit to extract genomic DNA, and then use the methods of Example 1 and Example 2 to perform typing detection of 7 genes. At the same time, the extracted DNA is genotyped and detected by the first-generation sequencing method, and the genotyping results of this method are 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 comparative test results of ACE (rs4646994) are shown in Table 8; ...

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Abstract

The invention relates to a detection composition, a kit and a detection method for guiding hypertension medication. According to the detection composition, the kit and the detection method, the genotypes of CYP2C9*3 (rs1057910), CYP2D6 (rs1065852), CYP3A5*3 (rs776746), ADRB1 (rs1801253), AGTR1 (rs5186), ACE (rs4646994) and NPPA (rs5065) are detected. The genotypes of the abovementioned seven sites are detected by utilizing the detection composition, the kit and the detection method, the method is simple and feasible, as well as rapid and efficient, the cost is low, and a novel path realizing simple and direct judgment is provided for the clinical medication of hypertension.

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 hypertension drug-related SNP sites. Background technique [0002] With the development of social economy and the change of residents' lifestyle, chronic non-communicable diseases (referred to as chronic diseases) have become a major public health problem affecting the health of residents in my country and even the world, and hypertension is one of the chronic diseases with a high prevalence rate. It is also the most important risk factor for cardiovascular and cerebrovascular diseases. According to the statistics of the World Health Organization (WHO), in 2012, the number of deaths from cardiovascular diseases in the world was 17 million, accounting for 46% of the deaths from chronic diseases, of which 9.4 million died from complications of hypertension, which has become the primary risk factor affect...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858C12Q1/6883C12Q2600/106C12Q2600/136C12Q2600/156C12Q2561/101
Inventor 林源吉丁佳女
Owner HANGZHOU KMB BIOTECH
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