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Method and kit for detecting rs2878677 locus of susceptibility gene of hypertension
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A kit and technology for hypertension, applied in the fields of molecular biology and medicine, can solve the problems of few studies and no proven correlation, and achieve the effects of low cost, simple diagnosis and treatment, and simple and easy method.
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 rs2878677 and EH
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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 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).
[0038] Primers for detection:
[0039] MFn2 gene rs2878677 upstream primer sequence: 5'-TTCTACTTTCTGTCCCTATGAATTTGA-3' (SEQ ID NO 2)
[0040] MFn2 gene rs2878677 downstream primer sequence: 5'-ACAAATACTGTATGATTCCACTTACGT-3' (SEQ ID NO 3) fluorescent probe:
[0041] MFn2 gene rs2878677 fluorescent probe 1: 5'-VIC-attctagaCgtctcacgt-TAMRA-3' (SEQ ID NO 4)
[0054] The rs2878677 site of the essential hypertension susceptibility gene MFn2 gene was detected by sequencing method. 10 cases of each of the above hypertension cases and control groups were selected for sequencing to determine the genotype of rs2878677.
[0055] 1. Experimental method
[0056] 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.
[0057] 2. Experimental results
[0058] In the end, the sequencing results of 20 cases were completely consistent with the genotype analysis results of 7900 fluorescent PCR.
[0059] 3. Association analysis of MFn2 gene rs2878677 genotype and hypertension susceptibility
[0060] Comparison of the distribution of MFn2 gene rs2878677 in hypertensive p...
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Abstract
The invention belongs to the field of molecular biology and medicine and relates to a method and a kit for detecting a susceptibility gene of hypertension. The invention provides a method for detecting a susceptibility gene of essential hypertension. The method comprises a step of detecting the gene type of a mitochondria fusion gene 2 (Mfn2) / hyperplasia suppressor gene (HSG) rs2878677 locus, wherein the hypertension susceptibility of an individual of which the rs2878677 locus carries a T gene type is obviously higher than that of common people. The invention also discloses a corresponding kit for detecting the susceptibility gene of hypertension. The kit comprises a primer for amplifying the rs2878677 locus and a primer for amplifying an area comprising the rs2878677 locus in the No.2 intron of Mfn2 gene. When used for detecting the gene type of the rs2878677 locus, the method is simple and is easy to implement, is fast and efficient, and has low cost and provides a simple and direct new way for the diagnosis and treatment of 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 rs2878677 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 温绍君李瑶刘阔李志忠吴海刘雅谢毅罗毅刘洁琳王佐广楼煜清严山文杰牛秋丽顾伟
Owner BEIJING ANZHEN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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