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  

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

作　　者：汪维鹏[1] 倪坤仪[2] 周国华[1] 

机构地区：[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

出　　处：《高等学校化学学报》2006年第10期1856-1858,共3页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:30270368)资助.

摘　　要：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.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 （ K281 del）, 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.

关 键 词：ALM-ASA 微流控芯片电泳 SNP CYP2D6 

分 类 号：O657[理学—分析化学]