Method for detecting hypertension susceptible gene and detection kit

A susceptibility gene 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

CN101892296AInactive Publication Date: 2010-11-24BEIJING ANZHEN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV +1
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2010-11-24
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention belongs to the field of molecular biology and medicine and relates to a method for detecting susceptibility of essential hypertension and a kit thereof. The method for detecting the susceptibility of the essential hypertension comprises a step of detecting genotypes of mitochondrion fusion gene 2 (Mfn2) / hyperplasia suppressor gene (HSG) rs4240897 sites of an individual. For an individual of which the rs4240897 carries a G genotype, the susceptibility of hypertension of is obviously higher than that of general people. The invention further discloses a corresponding detection kit which contains a primer of which a No.2 amplification Mfn2 gene comprises a sub-area. By using the invention to detect the genotypes of the rs4240897 sites, the method is simple and easy, has a high speed, high efficiency and a low cost, and provides a new simple approach for hypertension diagnosis and treatment.
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Description

technical field

[0001] The invention belongs to the fields of molecular biology and medicine. The invention relates to a method and a detection kit for detecting hypertension susceptibility genes. More specifically, it involves detection of the single nucleotide polymorphism (single nuclcotide polymorphism, SNP) site rs4240897 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 foot EH is still not completely clear, and the early diagnosis and prevention of hyperten...

Examples

Embodiment 1

[0048] Fluorescent PCR detection

[0049] 1. Experimental materials

[0050] The 7900HT fluorescent 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).

[0051] 2. Primer and probe design and synthesis

[0052] Using the partial sequence of intron 2 of the MFn2 gene as a template, the Primer ExpressTM2.0 software was used to analyze the TaqMan primers and probe sites, and custom-synthesized by Applied Biosystems (ABI).

[0053] Primers for detection:

[0054] MFn2 gene rs4240897 upstream primer sequence: 5′-GCTAGGAGTTGCTGTACATAAC-3′

[0055] MFn2 gene rs4240897 downstream primer sequence: 5′-AGGTGGTCAACAGCTTTGGCT-3′

[0056] Fluorescent probes:

[0057] MFn2 gene rs4240897 fluorescent probe 1: 5′-VIC-TACTCCTCAACAAATAG-TAMRA-3′

[0058] MFn2 gene rs4240897 fluorescent probe 2: 5′-FAM-TACTCCTCAACGAATAG-TAMRA-3′

[0059] 3. ...

Embodiment 2

[0073] The rs4240897 site of the essential hypertension susceptibility gene MFn2 gene was detected by sequencing. 10 cases of each of the above hypertension cases and control groups were selected for sequencing to determine the genotype of rs4240897.

[0074] 1. Experimental method

[0075] 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 the 3130xl genetic analyzer of ABI Company, which is analyzed with sequence analysis 5.2 analysis software, and the results can also be viewed with chromas.

[0076] 2. Experimental results

[0077] The screenshot of the sequencing result is as follows: figure 1 shown. In the end, the sequencing results of 20 cases were completely consistent with the genotype analysis results of 7900 fluorescent PCR.

[0078] 3. Association analysis of MFn2 gene rs4240897 genotype and hypertension susceptibility

...