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Method for detecting rs2295281 locus of hypertension susceptibility gene and detection kit
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A kit and site 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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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 rs2295281 and EH
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[0036] The 7900HT fluorescent quantitative PCR instrument was purchased from ABI Company of the United States, and the polymerasechain reaction liquid (TaqMan EXPress Master Mix) was custom-synthesized by Applied Biosystems (ABI).
[0037] 2. Primer and probe design and synthesis:
[0038] Using the partial sequence of intron 8 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).
[0039] Primers for detection:
[0040] MFn2 gene rs2295281 upstream primer sequence: 5'-CCACTTTGCTGGATGCAC-3' (SEQ ID NO 2)
[0041] MFn2 gene rs2295281 downstream primer sequence: 5'-AAGTCACCAGTAAGAACCAGT-3' (SEQ ID NO 3)
[0042] Fluorescent probes:
[0043] MFn2 gene rs2295281 fluorescent probe 1: 5'-VIC-CTTCAGTGCCGGTGCT-TAMRA-3' (SEQ ID NO 4)
[0057] The rs2295281 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 rs2295281.
[0058] 1. Experimental method
[0059] 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.
[0060] 2. Experimental results
[0061] In the end, the sequencing results of 20 cases were completely consistent with the genotype analysis results of 7900 fluorescent PCR.
[0062] 3. Association analysis of MFn2 gene rs2295281 genotype and hypertension susceptibility
[0063] Comparison of the distribution of MFn2 gene rs2295281 in hypertensiv...
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Abstract
The invention belongs to the field of molecular biology and medicine, relating to a method for detecting a hypertension susceptibility gene and a detection kit thereof. The method for detecting a primary hypertension susceptibility gene comprises the following step of: detecting the genotype of the rs2295281 locus of a mitochondrionfusion gene 2(Mfn2) / a hyperplasiasuppressorgene (HSG). The hypertension susceptibility of an rs2295281 individual with a genotype C is remarkably higher than that of the general public. The invention also discloses the corresponding detection kit. The detection kit contains a primer for amplifying the rs2295281 locus and can also contain the primer for amplifying a region comprising the rs2295281 locus in an eighth intron of the Mfn2 gene. The genotype of the rs2295281 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 involves detection of the single nucleotide polymorphism (single nucleotide polymorphism, SNP) site rs2295281 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...
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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 温绍君谢毅楼煜清李志忠吴海刘洁琳李瑶刘雅王佐广刘阔文杰严山
Owner BEIJING ANZHEN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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