连接反应介导的等位基因特异性扩增-微流控芯片电泳法同时检测多个SNP位点

高等学校化学学报

连接反应介导的等位基因特异性扩增-微流控芯片电泳法同时检测多个SNP位点

汪维鹏^1,2, 倪坤仪², 周国华^1,2

1. 华东医学生物技术研究所, 南京 210002;
2. 中国药科大学分析化学教研室, 南京 210038

收稿日期:2006-01-10 修回日期:1900-01-01 出版日期:2006-10-10 发布日期:2006-10-10
通讯作者: 周国华

Simultaneous Detection of Multiplex SNPs by Adapter-ligation Mediated Allele-specific Amplification(ALM\|ASA) on CE-chip Electrophoresis Device

WANG Wei-Peng^1,2, NI Kun-Yi², ZHOU Guo-Hua^1,2

1. Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002, China;
2. Department of Analytical Chemistry, China Pharmaceutical University, Nanjing 210038, China

Received:2006-01-10 Revised:1900-01-01 Online:2006-10-10 Published:2006-10-10
Contact: ZHOU Guo-Hua

摘要/Abstract

摘要： 以CYP2D6基因中的6个SNP位点为测定对象, 开展多个SNP位点同时测定的方法学研究.

关键词: ALM-ASA, 微流控芯片电泳, SNP, CYP2D6

Abstract: To detect multiplex single nucleotide polymorphisms(SNPs) simultaneously, a new method was established by combining ALM-ASA with microfabricated CE-chip. Taking the CYP2D6 gene as an example, six SNPs, 100C>T(P34S), 1707T>del(frameshift), 1758G>T(stop codon), 2470T>C, 2549A>del(frameshift) and 2613AGA>del(K281del), were typed by four steps consisting of preamplification, digestion and ligation, allele-specific amplification, and amplicon separation by chip-CE. The genotyping results of 20 different genome samples by 6-plexed ALM-ASA were completely consistent with those obtained by polymerase chain reaction-restriction fragment length polymorphism analysis(PCR-RFLP), indicating that the multiplex approach established in this study was accurate and inexpensive. As the small reagent consumption by CE-chip device, a low cost for SNP typing was achieved together with the multiplex PCR technology proposed in this report. Neither modification of microchip channels nor clean-up process of PCR products was required; this greatly shortens the whole time for SNP typing.

Key words: ALM-ASA, Microchip electrophoresis, SNP, CYP2D6

中图分类号:

O657

TrendMD:

汪维鹏,倪坤仪,周国华 . 连接反应介导的等位基因特异性扩增-微流控芯片电泳法同时检测多个SNP位点. 高等学校化学学报, doi: .

WANG Wei-Peng^1,2, NI Kun-Yi², ZHOU Guo-Hua^1,2. Simultaneous Detection of Multiplex SNPs by Adapter-ligation Mediated Allele-specific Amplification(ALM\|ASA) on CE-chip Electrophoresis Device. Chem. J. Chinese Universities, doi: .