一种同时检测多个单核苷酸多态性位点的新方法  

Simultaneous Detection of Several Single Nucleotide Polymorphisms

作　　者：赵春霞[1] 石先哲[1] 张岩[2] 马坚妹[2] 牟红梅[1] 吕申[1] 许国旺[1] 

机构地区：[1]中国科学院大连化学物理研究所国家色谱研究分析中心,大连116011 [2]大连医科大学第二附属医院细胞实验室,大连116023

出　　处：《分析化学》2003年第8期906-910,共5页Chinese Journal of Analytical Chemistry

基　　金：中国科学院知识创新工程前沿资助项目 (K2 0 0 1A4)

摘　　要：对人类基因组中的单核苷酸多态性进行快速而准确的定位和分型是人类基因组计划的重要内容。本文利用SNaPshot试剂盒 ,建立了一种以多重引物延伸为基础、可同时对多个已知单核苷酸多态性位点进行快速而准确的遗传分型的方法。利用这一方法检测了 2 0例大肠癌病人肿瘤组织中错配修复基因hMLH1和抑癌基因P5 3的 8个单核苷酸多态性位点 (包括 5个突变热点 ) ,其中一例发现错配修复基因hMLH1的 384位密码子处发生GTT→GAT的杂合性突变。对该基因外显子的直接测序结果与SNaPshot检测结果完全吻合 ,充分证明了该方法的可靠性。Accurate and fast genotyping of single nucleotide polymorphisms (SNPs) is of significant scientific importance in human genome project. Here a new automated fluorescent method that can rapidly and accurately genotype multiplex known SNPs was developed, which was achieved by substantially improving a commercially available protocol. By this method, mismatch repair gene hMLH1 and cancer repression gene P53 were investigated, eight known SNP spots (including five hot spots) of 20 coloeretal cancer patients' hMLH1 and P53 gene were detected, and a heterozygosity mutation (GTT-->GAT) at codon 384 of hMLH1 was found. Sequencing result of hMLH1 proves the reliability of this new method.

关 键 词：同时检测 单核苷酸 多态性 基因组 肿瘤检测 产物纯化 DNA序列 病因学 大肠癌 错配修复基因hMLHl P53基因 

分 类 号：Q52[生物学—生物化学]