AS-PCR检测ABCG2 34G>A多态性

中国临床药理学与治疗学 ›› 2014, Vol. 19 ›› Issue (10): 1111-1115.

AS-PCR检测ABCG2 34G>A多态性

李泰霖¹, 谢海棠³, 许标波¹, 彭艳¹, 万子睿¹, 孙红¹, 曾樱¹, 沈杰³, 王果¹, 朱院山^{1, 2}

¹ 中南大学临床药理研究所,长沙 410078,湖南;
² Department of Medicine of Weill Cornell Medical College,New York,NY 10065;
³ 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,芜湖 241001,安徽

收稿日期:2014-03-22 修回日期:2014-09-26 出版日期:2014-10-26 发布日期:2014-10-29
通讯作者: 王果,男,副教授,主要研究方向为遗传药理学与临床药理学。Tel: 0731-84805380 E-mail: 1326441727@qq.com
作者简介:李泰霖,男,硕士研究生,主要研究方向为遗传药理学。 Tel: 15273229024 E-mail: 416514282@qq.com
基金资助:
国家自然科学基金项目(81072706,81173134); 湖南省科技计划项目(2013FJ3036); 安徽省自然科学基金项目(1408085MH163)

Genotyping of ABCG2 34G>A polymorphism by allele-specific PCR

LI Tai-lin¹, XIE Hai-tang³, XU Biao-bo¹, PENG Yan¹, WAN Zi-rui¹, SUN Hong¹, ZENG Ying¹, SHEN Jie³, WANG Guo¹, ZHU Yuan-shan²

¹ Institute of Clinical Pharmacology, Central South University, Changsha 410078, Hunan,China;
² Department of Medicine of Weill Cornell Medical College,New York,NY 10065;
³ Institute of Clinical Pharmacy and Pharmacology, Yijishan Hospital of Wannan Medical College, Wuhu 241001, Anhui, China

Received:2014-03-22 Revised:2014-09-26 Online:2014-10-26 Published:2014-10-29

摘要/Abstract

摘要： 目的建立和优化检测ABCG2 34G>A多态性的等位基因特异性PCR方法(allele-specific PCR,AS-PCR),并对100例健康受试者进行分型检测。方法设计合成3'-末端错配引物和包含内部错配位点的3'-末端错配引物,进行AS-PCR扩增,比较两者的扩增特异性,应用AS-PCR检测100例健康受试者ABCG2 34位多态型,采用焦磷酸测序方法(pyrosequencing)进行抽样验证。结果含有第3位内部错配位点的3'-末端错配引物扩增特异性明显优于含有第2位内部错配位点以及不含内部错配位点的3'-末端错配引物。100例样本AS-PCR分型结果与同批标本前期的焦磷酸测序结果一致。ABCG2 34AA,34GA和34GG型频率分别为7%、32%、61%。结论本文所建立的AS-PCR方法在进行ABCG2 34位多态分型时,具有省时、快速和成本低等优点,经过在引物中引入内部错配位点等优化设计后,分型结果准确可靠,适合临床应用。

关键词: 等位基因特异性PCR, ABCG2, 单核苷酸多态性, BCRP

Abstract: AIM: To establish an allele-specific PCR method to genotype ABCG2 34 G>A polymorphism and verify it using 100 healthy subjects gDNA samples. METHODS: Allele-specific PCR primers with or without an additional mismatch nucleotide artificially introduced within the two bases closest to the 3' end were designed, and with different primer pairs, AS-PCR was used to detect ABCG2 34 allele of 100 gDNA samples. The Genotyping results were compared to pyrosequencing results of the same samples we did before. RESULTS: The discrimination ability of 34F/34Rn2 primer pairs was significantly better than the others, and the frequency of ABCG2 34AA, 34GA and 34GG of the 100 gDNA samples were 7%, 32% and 61% respectively, which was consistent with pyrosequencing. CONCLUSION: The results suggest the discrimination ability of AS-PCR method established in this study has been greatly improved and that will facilitate this time-saving, rapid and inexpensive technique into clinical application.

Key words: allele-specific PCR, ABCG2, single nucleotide polymorphism, BCRP

中图分类号:

R968

李泰霖, 谢海棠, 许标波, 彭艳, 万子睿, 孙红, 曾樱, 沈杰, 王果, 朱院山. AS-PCR检测ABCG2 34G>A多态性[J]. 中国临床药理学与治疗学, 2014, 19(10): 1111-1115.

LI Tai-lin, XIE Hai-tang, XU Biao-bo, PENG Yan, WAN Zi-rui, SUN Hong, ZENG Ying, SHEN Jie, WANG Guo, ZHU Yuan-shan. Genotyping of ABCG2 34G>A polymorphism by allele-specific PCR[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2014, 19(10): 1111-1115.

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