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Method for detecting mononucleotide polymorphism locus rs2236058 of hypertension susceptibility genes and detection kit

A kit and high blood pressure technology, applied in the fields of molecular biology and medicine, can solve the problems of less research and unconfirmed correlation, and achieve the effects of low cost, simple diagnosis and treatment, and simple and easy methods

Inactive Publication Date: 2012-11-21
BEIJING ANZHEN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV +1
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  • Summary
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  • 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 rs2236058 and EH

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  • Method for detecting mononucleotide polymorphism locus rs2236058 of hypertension susceptibility genes and detection kit
  • Method for detecting mononucleotide polymorphism locus rs2236058 of hypertension susceptibility genes and detection kit
  • Method for detecting mononucleotide polymorphism locus rs2236058 of hypertension susceptibility genes and detection kit

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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 11 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 rs2236058 upstream primer sequence: 5'-CTTGGGCAGTTAGGACACGC-3' (SEQ ID NO 2)

[0040] MFn2 gene rs2236058 downstream primer sequence: 5'-CCTCCAACACAGGGAATCG-3' (SEQ ID NO 3)

[0041] Fluorescent probes:

[0042] MFn2 gene rs2236058 fluorescent probe 1: 5'-VIC-TCATCACCCCACCTGGTC-TAMRA-3' (SEQ ID NO4)

[0043] MFn2 gene rs2236058 fluorescent prob...

Embodiment 2

[0055] The rs2236058 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 rs2236058.

[0056] 1. Experimental method

[0057] 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.

[0058] 2. Experimental results

[0059] The screenshot of the sequencing result is as follows: figure 2 shown.

[0060] In the end, the sequencing results of 20 cases were completely consistent with the genotype analysis results of 7900 fluorescent PCR.

[0061] 3. Association analysis of MFn2 gene rs2236058 genotype and hypertension susceptibi...

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Abstract

The invention belongs to the fields of molecular biology and medicine and relates to a method for detecting hypertension susceptibility genes and a detection kit thereof. The method for detecting the essential hypertension susceptibility genes comprises the step of detecting the gene type of the locus rs2236058 of mitochondria fusion genes 2 (Mfn2) / hyperplasia suppressor genes (HSG). People having the locus rs2236058 of the C gene type are more susceptible to the hypertension than the common crowd. The invention also discloses a relevant detection kit containing primers for amplifying the locus rs2236058 and primers for amplifying a zone containing the locus rs2236058 in the intron No.11 of the Mfn2 genes. The method is easy to operate, can be used for quickly and effectively detecting the gene type of the locus rs2236058 and has low cost, thereby being a simple new method for diagnosing and treating the hypertension.

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 rs2236058 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 Patents(China)
IPC IPC(8): C12Q1/68
Inventor 温绍君李瑶吴海李志忠谢毅刘雅刘洁琳王佐广楼煜清刘阔严山顾伟牛秋丽文杰
Owner BEIJING ANZHEN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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