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Method for detecting hypertension susceptibility gene and detection kit
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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
Inactive Publication Date: 2013-06-12
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 rs1474868 and EH
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Embodiment 1
[0028] Embodiment 1 Fluorescence PCR detects
[0029] 1. Experimental materials
[0030] 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).
[0031] 2. Primer and probe design and synthesis:
[0032] 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).
[0033] Primers for detection:
[0034] MFn2 gene rs 1474868 upstream primer sequence: 5'-TGAACTCCCAGCAGACCTTGTA-3' (SEQ ID NO 2)
[0035] MFn2 gene rs1474868 downstream primer sequence: 5'-GGTGCTTTTAAGTTCTGGTCAATGT-3' (SEQ ID NO 3)
[0036] Fluorescent probes:
[0037] MFn2 gene rs1474868 fluorescent probe 1: 5'-VIC-TGCAAGACTCGACAGC-TAMRA-3' (SEQ ID NO 4)
[0038] MFn2 gene rs1474868 fluorescent pro...
Embodiment 2
[0049] The rs1474868 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 rs1474868.
[0050] 1. Experimental method
[0051] 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.
[0052] 2. Experimental results
[0053] In the end, the sequencing results of 20 cases were completely consistent with the genotype analysis results of 7900 fluorescent PCR.
[0054] 3. Association analysis of MFn2 gene rs1474868 genotype and hypertension susceptibility
[0055] Comparison of the distribution of MFn2 gene rs1474868 in hypertensive p...
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Abstract
The invention belongs to the fields of molecular biology and medical science, relates to a method for detecting a hypertension susceptibility gene and a detection kit thereof and provides a method for detecting a primary hypertension susceptibility gene. The method comprises the following step of: detecting the genotype of a mitochondrial fusion gene 2 (Mfn2) / hyperplasic suppress gene (HSG) at anrs1474868 locus, wherein the hypertension susceptibility of an individual with a G type gene at the rs1474868 is remarkably higher than that of ordinary people. The invention also discloses a corresponding detection kit. The detection kit contains a primer for amplifying the rs1474868 locus and can also comprise a primer for amplifying an area which contains the rs1474868 locus in a second intronof the Mfn2 gene. The genotype of the rs1474868 locus is detected by the method. The method has the advantages of simpleness, practicability, rapidness, high efficiency and low cost, and a new simpleand convenient method is provided 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 rs1474868 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 温绍君谢毅王佐广李志忠吴海李瑶罗毅刘雅刘洁琳楼煜清刘阔柳景华
Owner BEIJING ANZHEN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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