Method for detecting mononucleotide polymorphism rs2236055 locus of hypertension susceptibility gene and kit thereof

A technology for single nucleotide polymorphism and hypertension, applied in the fields of molecular biology and medicine, can solve the problems of unproven correlation and little research, and achieve the effects of simple diagnosis and treatment, low cost, and simple and easy method.

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 rs2236055 and EH

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  • Method for detecting mononucleotide polymorphism rs2236055 locus of hypertension susceptibility gene and kit thereof
  • Method for detecting mononucleotide polymorphism rs2236055 locus of hypertension susceptibility gene and kit thereof
  • Method for detecting mononucleotide polymorphism rs2236055 locus of hypertension susceptibility gene and kit thereof

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Embodiment 1

[0044] Fluorescent PCR detection

[0045] 1. Experimental materials

[0046] The 7900HT fluorescence quantitative PCR instrument was purchased from ABI Company in the United States, and the polymerase chain reaction solution (TaqManEXPress Master Mix) was custom-synthesized by Applied Biosystems (ABI).

[0047] 2. Primer and probe design and synthesis:

[0048] Using the partial sequence of intron 2 of the MFn2 gene as a template, TaqMan primers and probe sites were analyzed using Primer ExpressTM 2.0 software, and custom-synthesized by Applied Biosystems (ABI).

[0049] Primers for detection:

[0050] MFn2 gene rs2236055 upstream primer sequence: 5′-TGTGCTCTCCTGGACCTCTG-3′

[0051] MFn2 gene rs2236055 downstream primer sequence: 5′-GAAGAGCCAGCACAGCAGAGA-3′

[0052] Fluorescent probes:

[0053] MFn2 gene rs2236055 fluorescent probe 1: 5′-VIC-CAAGTCTCTTGAATG-TAMRA-3′

[0054] MFn2 gene rs2236055 fluorescent probe 2: 5′-FAM-CAAGTCTCTTGGATG-TAMRA-3′

[0055] 3. Sample test...

Embodiment 2

[0069] The rs2236055 site of the essential hypertension susceptibility gene MFn2 gene was detected by sequencing. 10 cases of each of the above-mentioned hypertension cases and control groups were selected for sequencing to determine the genotype of rs2236055.

[0070] 1. Experimental method

[0071] 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.

[0072] 2. Experimental results

[0073] In the end, the sequencing results of 20 cases were completely consistent with the genotype analysis results of 7900 fluorescent PCR.

[0074] 3. Association analysis of MFn2 gene rs2236055 genotype and susceptibility to hypertension

[0075] RxCχ2 test was used to compare the distribution of MFn2 gene rs22...

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Abstract

The invention belongs to the field of molecular biology and medicine, relating to a method for detecting primary hypertension susceptibility and a kit thereof. The method for detecting primary hypertension susceptibility comprises the following step of: detecting the genotype of the rs2236055 locus of a mitochondrion fusion gene 2(Mfn2)/a hyperplasia suppressor gene (HSG) of an individual. The hypertension susceptibility of the rs2236055 individual with an genotype A is remarkably higher than that of the general public. The invention also discloses the corresponding detection kit. The kit contains a primer for amplifying a primer of a region in the second intron of the Mfn2 gene. The genotype of the rs2236055 locus is detected by adopting the method. The invention has simple and easy method, quick speed and high efficiency and low cost and provides a novel shortcut approach for diagnosing and treating hypertension.

Description

technical field [0001] The present invention relates to the fields of molecular biology and medicine. More specifically, it relates to the detection of the single nucleotide polymorphism (single nucleotide polymorphism, SNP) site rs2236055 of human mitochondrial fusion gene 2 (Mfn2 gene) and 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 the environment,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 温绍君谢毅李志忠吴海王佐广李瑶罗毅刘洁琳刘雅楼煜清刘阔柳景华
Owner BEIJING ANZHEN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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