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Detection method of single nucleotide polymorphism sites of hypertension susceptibility gene and kit thereof

A single nucleotide polymorphism and high blood pressure technology, applied in the fields of molecular biology and medicine, can solve the problems of unconfirmed correlation and few studies, and achieve the effects of simple diagnosis and treatment, low cost, and simple and easy methods

Inactive Publication Date: 2013-02-27
BEIJING ANZHEN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 rs3753579 and EH

Method used

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  • Detection method of single nucleotide polymorphism sites of hypertension susceptibility gene and kit thereof
  • Detection method of single nucleotide polymorphism sites of hypertension susceptibility gene and kit thereof
  • Detection method of single nucleotide polymorphism sites of hypertension susceptibility gene and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Fluorescent PCR detection

[0043] 1. Experimental materials

[0044] The 7900HT fluorescence quantitative PCR instrument was purchased from ABI Company of the United States, and the polymerase chain reaction solution (TaqMan EXPressMaster Mix) was custom-synthesized by Applied Biosystems (ABI).

[0045] 2. Primer and probe design and synthesis:

[0046] Using the partial sequence of the 5' promoter region of the MFn2 gene as a template, the TaqMan primers and probe sites were analyzed using Primer ExpressTM 2.0 software, and custom-synthesized by Applied Biosystems (ABI).

[0047] Primers for detection:

[0048] MFn2 gene rs3753579 upstream primer sequence: 5′-GGAGGAGGAAACAATCCGAC-3′

[0049] MFn2 gene rs3753579 downstream primer sequence: 5′-GGTTTATCAGTCTCATGACGATG-3′

[0050] Fluorescent probes:

[0051] MFn2 gene rs3753579 fluorescent probe 1: 5′-VIC-ATGTAGCGTGCCTTG-TAMRA-3′

[0052] MFn2 gene rs3753579 fluorescent probe 2: 5′-FAM-ATGTAGCGTGCTTTG-TAM...

Embodiment 2

[0067] The rs3753579 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 rs3753579.

[0068] 1. Experimental method

[0069] 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.

[0070] 2. Experimental results

[0071] The screenshot of the sequencing results showed that, in the end, the sequencing results of 20 cases were completely consistent with the genotype analysis results of 7900 fluorescent PCR.

[0072] 3. Association analysis of MFn2 gene rs3753579 genotype and hypertension susceptibility

[0073] Comparison of ...

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Abstract

The invention relates to the molecular biology and medicine field, in particular to a method for detecting essential hypertension susceptibility, which comprises the following steps of detecting the genotype at a site of rs3753579 of an individual mitochondrion fusion gene 2 (Mfn2) / hyperplasia suppressor gene (HSG), and thus judging whether the risk of the individual to suffer from essential hypertension is larger than that of common people. The invention also discloses a corresponding detection kit. By detecting the genotype at the site of rs3753579, the method is simple, practicable, fast, high efficient and low-cost, and provides a new short-cut way to the diagnosis and the treatment of 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 rs3753579 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 Patents(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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