焦磷酸测序技术检测SLCO1B1基因多态性方法的建立  

Development of pyrosequencing method for detection of SLCO1B1 polymorphisms

作　　者：万子睿[1] 谢海棠[2] 郭栋[1] 胡东莉[1] 王丽萍[1] 王果[1] 

机构地区：[1]中南大学临床药理研究所,湖南长沙410078 [2]皖南医学院弋矶山医院临床药学部安徽省药物临床评价中心,安徽芜湖241001

出　　处：《中国临床药理学与治疗学》2012年第5期543-548,共6页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金项目(81072706;81173134);高等学校博士学科点专项科研基金资助课题(20090162120024);湖南省科技计划项目(2009JT3020)

摘　　要：目的:建立SLCO1B1A388G和T521C单核苷酸多态位点的焦磷酸测序方法,分析中国健康人群中分布频率。方法:制备300例健康人外周血gDNA,应用PyroMark ID焦磷酸测序仪进行多态位点分型分析。并通过重复性检验和毛细管电泳测序验证方法正确性。结果:建立了SL-CO1B1A388G和T521C多态性分析的焦磷酸测序新方法,经毛细管电泳测序验证和重复性验证,结果准确可靠。在300例标本中,388A、388G、521T、521C等位基因频率分别为28%、72%、89.5%和10.5%,符合Hardy-Weinberg平衡。结论:焦磷酸测序方法可准确、高通量、快速检测SLCO1B1A388G和T521C单核苷酸多态性,特别适宜大样本量的临床及科研批量检测需要。To establish a pyrosequenc- ing based method for detection SLCOIB1 A388G and T521C polymorphisms and to determine the frequency of these polymorphisms in healthy Chinese. METHODS： After preparation of gD- NA from blood of 300 subjects, the target frag-ments were amplified by PCR, polymorphisms were detected on PyroMark ID by pyrosequenc- ing technology. The reliability of pyrosequenc- ing methods were validated by repeat tests and Sanger sequencing. RESULTS. We established a new pyrosequencing method to detect the SL-CO1B1 A388G and T521C polymorphisms poly- morphisms in healthy Chinese. The detection rate and repetition rate were both 100%. The frequencies of 388A, 388G, 521T and 521C al- leles were 28%0,72%o,89. 5G and 10. 5G, re- spectively. Genotype frequencies match the Har- dy-Weinberg equilibrium. CONCLUSION. These pyrosequencing assays to detect SLCO1B1 poly-morphisms are proved to be a rapid, accurate and high-throughput alternative to conventional methods, and it can be a preferred option in re- search and clinical application.

关 键 词：焦磷酸测序 SLCO1B1 OATP1B1 单核苷酸多态性 

分 类 号：R968[医药卫生—药理学]