Detection method and kit of single nucleotide polymorphism locus rs7550536 of susceptibility gene of hypertension

A detection method, high blood pressure technology, applied in the fields of molecular biology and medicine, can solve the problems of less research and no proven correlation, and achieve the effects of low cost, simple diagnosis and treatment, and simple and easy methods

Inactive Publication Date: 2010-11-24
BEIJING ANZHEN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV +1
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Problems solved by technology

Therefore, the HSG gene is one of the candidate genes for essential hypertension. Later gene function research shows that the HSG gene is involved in mitochondrial fusion, so it is officially named mitochondrial fusion gene 2 (mitofusin 2, Mfn2). However, the current research on Mfn2 and EH Fewer, no reports confirming the association between rs7550536 and EH

Method used

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  • Detection method and kit of single nucleotide polymorphism locus rs7550536 of susceptibility gene of hypertension
  • Detection method and kit of single nucleotide polymorphism locus rs7550536 of susceptibility gene of hypertension
  • Detection method and kit of single nucleotide polymorphism locus rs7550536 of susceptibility gene of hypertension

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Embodiment 1

[0033] Embodiment 1 Fluorescence PCR detects

[0034] 1. Experimental materials

[0035] The 7900HT fluorescent quantitative PCR instrument was purchased from ABI Company of the United States, and the polymerase chain reaction liquid (TaqMan EXPress Master Mix) was custom-synthesized by Applied Biosystems (ABI).

[0036] 2. Primer and probe design and synthesis:

[0037] Using the partial sequence of intron 14 of the MFn2 gene as a template, the Primer ExpressTM 2.0 software was used to analyze the TaqMan primers and probe sites, and custom-synthesized by Applied Biosystems (ABI).

[0038] Primers for detection:

[0039] MFn2 gene rs7550536 upstream primer sequence: 5'-AACATGAAGGCTCCTTGGCT-3' (SEQ ID NO 2)

[0040] MFn2 gene rs7550536 downstream primer sequence: 5'-ACACCTGCAGAGCACATGGT-3' (SEQ ID NO 3) fluorescent probe:

[0041] MFn2 gene rs7550536 fluorescent probe 1: 5'-VIC-CCTGGTAGGGATGGG-TAMRA-3' (SEQ ID NO 4)

[0042] MFn2 gene rs7550536 fluorescent probe 2: 5'-FAM-...

Embodiment 2

[0054] The rs7550536 site of the essential hypertension susceptibility gene MFn2 gene was detected by sequencing method. 10 cases of the above-mentioned hypertension case-control samples were selected for sequencing to determine the genotype of rs7550536.

[0055] 1. Experimental method

[0056] The above-mentioned fluorescent PCR primers were still used as primers for PCR sequencing, and the amplified products were directly sequenced after purification. The sequencing instrument is ABI's 3130xl genetic analyzer, which is analyzed with sequence analysis 5.2 analysis software, and the results can also be viewed with chromas.

[0057] 2. Experimental results

[0058] The screenshot of the sequencing result is as follows: figure 2 shown.

[0059] In the end, the sequencing results of 20 cases were completely consistent with the genotype analysis results of 7900 fluorescent PCR.

[0060] 3. Association analysis of MFn2 gene rs7550536 genotype and hypertension susceptibility ...

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Abstract

The invention belongs to the molecular biology and medicine field and relates to a method for detecting a susceptibility gene of hypertension and a detection kit thereof. The invention provides a method for detecting a susceptibility gene of essential hypertension, comprising the step of detecting the genetype of a mitochondrion fusion gene 2(Mfn2) / hyperplasia suppressor gene (HSG) locus rs7550536. The individual, rs7550536 of which carries the T genetype has obviously higher susceptibility of hypertension than the common people. The invention also discloses the corresponding detection kit which contains a primer amplifying the locus rs7550536 and a primer amplifying the region of the locus rs7550536 contained in the No.14 intron of the Mfn2 gene. Being used for detecting the genetype of the locus rs7550536, the method is simple and practicable, fast and efficient and low in cost and provides a simple and direct new way for hypertension diagnosis and treatment.

Description

technical field [0001] The present invention relates to the fields of molecular biology and medicine. More specifically, it involves detection of the single nucleotide polymorphism (single nucleotide polymorphism, SNP) site rs7550536 of human mitochondrial fusion gene 2 (Mfn2 gene) and its correlation with essential hypertension. The invention also relates to a method and a kit for detecting the SNP site. Background technique [0002] Essential hypertension (essential hypertension, EH) is a multifactorial and polygenic disease. It is a common and frequent cardiovascular disease caused by environmental and genetic factors, which has a great impact on human health. With the development of molecular medicine, more than 150 hypertension-related genes have been discovered, but the pathogenesis of EH is still not completely clear, and the early diagnosis and prevention of hypertension have not been fully resolved. EH is the result of the combined effects of genetic factors and t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 温绍君谢毅柳景华李志忠李瑶刘洁琳刘雅吴海王佐广楼煜清刘阔严山牛秋丽
Owner BEIJING ANZHEN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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