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Development of a high resolution melting method for genotyping of risk HLA-DQA1 and PLA2R1 alleles and ethnic distribution of these risk alleles

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单位: [1]Huazhong Univ Sci & Technol,Tongji Med Coll,Tongji Hosp,Inst Hypertens,Wuhan 430030,Peoples R China [2]Huazhong Univ Sci & Technol,Tongji Med Coll,Tongji Hosp,Dept Internal Med,Wuhan 430030,Peoples R China [3]Huazhong Univ Sci & Technol,Tongji Med Coll,Tongji Hosp,Genet Diag Ctr,Wuhan 430030,Peoples R China [4]Univ Laval, Ctr Rech, Inst Univ Cardiol & Pneumol Quebec, Quebec City, PQ, Canada
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关键词: Idiopathic membranous nephropathy HLA-DQA1 PLA2R1 High resolution melting HRM

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Recent studies have demonstrated that alleles at single nucleotide polymorphisms (SNPs) rs2187668 and rs4664308 within genes HLA-DQA1 and PLA2R1, respectively, had a significant impact on the susceptibility to idiopathic membranous nephropathy (IMN). Analysis of the two genomic loci could identify alleles for individuals at risk for IMN. Conventional methods for genotyping are labor intensive, expensive or time consuming. High resolution melting (HRM) is a new technique for genotyping and has the advantages of simplicity, speed, high sensitivity and low cost. Here, we describe genotyping of SNPs rs2187668 and rs4664308 using HRM. In this study, we identified polymorphisms of rs2187668 and rs4664308 in 480 healthy unrelated Chinese volunteers of two ethnic groups from three different geographical areas in China. The two genomic loci were genotyped by HRM using a saturating fluorescent dye SYTO (R) 9 on 7900 HT and RG 6000 instruments, and were further confirmed by direct DNA sequencing. Three different SNP genotypes were sufficiently distinguished by HRM with mean sensitivity of 98.8% and mean error rate of 1.9%. In addition, the allele frequencies varied greatly based on ethnic or geographic origins. In conclusion, HRM is a rapid, cost efficient, sensitive, suitable technique for genotyping, and simple enough to be readily implemented in a diagnostic laboratory. We believe this will be a valuable technique for determining the genotype of rs2187668 and rs4664308 and for assessing individual susceptibility to IMN. (C) 2012 Elsevier B.V. All rights reserved.

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出版当年[2012]版:
大类 | 3 区 生物
小类 | 4 区 遗传学
最新[2025]版:
大类 | 3 区 生物学
小类 | 3 区 遗传学
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出版当年[2011]版:
Q3 GENETICS & HEREDITY
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Q2 GENETICS & HEREDITY

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第一作者单位: [1]Huazhong Univ Sci & Technol,Tongji Med Coll,Tongji Hosp,Inst Hypertens,Wuhan 430030,Peoples R China [2]Huazhong Univ Sci & Technol,Tongji Med Coll,Tongji Hosp,Dept Internal Med,Wuhan 430030,Peoples R China [3]Huazhong Univ Sci & Technol,Tongji Med Coll,Tongji Hosp,Genet Diag Ctr,Wuhan 430030,Peoples R China
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通讯机构: [1]Huazhong Univ Sci & Technol,Tongji Med Coll,Tongji Hosp,Inst Hypertens,Wuhan 430030,Peoples R China [2]Huazhong Univ Sci & Technol,Tongji Med Coll,Tongji Hosp,Dept Internal Med,Wuhan 430030,Peoples R China [3]Huazhong Univ Sci & Technol,Tongji Med Coll,Tongji Hosp,Genet Diag Ctr,Wuhan 430030,Peoples R China [*1]Huazhong Univ Sci & Technol,Tongji Med Coll,Tongji Hosp,Inst Hypertens,1095 Jiefang Ave,Wuhan 430030,Peoples R China
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