Rapid genotyping of APOA5 鈭?#xa0;1131T>C polymorphism using high resolution melting analysis with unlabeled probes

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• 作者：Song-mei Liu^a ; ^b ; ^{liu_sm@126.com} ; Feng-xia Xu^a ; Fan Shen^a ; Yan Xie^a
• 关键词：ApoA5 ; apolipoprotein A5 ; PCR ; polymerase chain reaction ; HRM ; high resolution melting ; SNP ; single nucleotide polymorphism ; Tm ; melting temperature
• 刊名：Gene
• 出版年：2012
• 期刊代码：73_03781119
• 类别：bio
• 出版时间：1 May, 2012
• 卷：498
• 期：2
• 页码：276-279
• 文件大小：663 K

摘要

The APOA5 鈭?#xA0;1131 T/C polymorphism (rs662799) exhibits a very strong association with elevated TG levels in different racial groups. High resolution melting (HRM) analysis with the use of unlabeled probes has shown to be a convenient and reliable tool to genotyping, but not yet been used for detecting rs662799 polymorphism. We applied the unlabeled probe HRM analysis and direct DNA sequencing to assay the 鈭?#xA0;1131T>C SNP in 130 cases DNA samples blindly. This HRM analysis can be completed in < 3 min for each sample. The two melting peaks were displayed at 66.1 卤 0.4 掳C for CC homozygote and 68.7 卤 0.2 掳C for TT homozygote; TC heterozygote showed the both melting peaks. The genotyping results by HRM method were completely concordant with direct DNA sequencing. The distribution of CC, TC, and TT genotypes for the 鈭?#xA0;1131T>C SNP was 9.2, 49.2, and 41.5%, respectively. This assay was sensitive enough to detect C allele down to 20%and 10%for T allele. The limit of detection for C and T allele was 6.2 and 2.5 ng/渭L DNA, respectively. The developed unlabeled probe HRM method provides an alternative mean to detect ApoA5 鈭?#xA0;1131T>C SNP rapidly and accurately.