冠心病相关基因多重单核苷酸多态性分型检测法的构建

摘要

Objective 　To establish a multiplex genotyping method for single nucleotide polymorphism s （SNPs） of genes associated with coronary artery disease （CAD ） .Methods 　DNA samples were extracted from peripheral white blood cells and 44 tag SNPs of genes associated with CAD for Chinese Han nationality were selected in the current work .Three primers were designed for each SNP .After polymerase chain reaction ，extension and hybridization ，genotypes of SNPs were investigated with the Genome Lab SNP stream genotyping system .Results 　Thirty two SNPs were genotyped and the rate of succeed SNP genotyping w as 72 .7％ （32/44） .The average genotyped rate of the 32 SNPs was 98 .3％ in the test of 583 DNA samples .Conclusion 　Thirty two plexs SNPs genotyping methods established in the current work were quick ，exact ，high-throughput ，easy operation and m icroanalyse .%目的 构建检测冠心病(CAD)相关基因多重单核苷酸多态性(SNP)分型的方法.方法 提取外周血白细胞DNA,选择CAD相关基因中国汉族人群的44个标签SNP,每个SNP设计3条引物,通过PCR扩增、延伸及杂交,应用SNP Stream基因分型系统对SNP位点进行基因分型.结果 构建的44个SNP分型成功32个,分型成功率为72.7%(32/44).通过对583份DNA标本进行检测,32个SNP的平均样本检出率为98.3%.结论 本实验构建的32重SNP基因分型检测方法具有快速、准确、高通量、操作简单、微量化等优点.