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ntPCR-RFLP检测HBV阿德福韦耐药变异—rtA181V变异
http://www.100md.com 闫 杰, 谢 雯, 王 磊, 王晶波, 冯 鑫,
阿德福韦;肝炎病毒;乙型;变异,闫杰,谢雯,冯鑫,宋淑静,刘顺爱,魏红山,王磊,王晶波,通讯作者:,Applicationofn
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     闫杰, 谢雯, 冯鑫, 宋淑静, 刘顺爱, 魏红山, 北京地坛医院 北京市 100011

    王磊, 王晶波, 山东大学医学院济南市传染病医院 山东省济南市 250021

    首都医学发展科研基金, No.2002-3046

    通讯作者: 闫杰, 100011, 北京市安外大街地坛公园13号, 北京地坛医院. jieyan@bbn.cn

    电话: 010-64211031-2330 传真:010-64281540

    收稿日期: 2006-02-06 接受日期: 2006-02-16

    Application of nest PCR-RFLP in the detection of adefovir dipivoxil resistance-associated mutation in hepatitis B virus

    Jie Yan, Wen Xie, Lei Wang, Jing-Bo Wang, Xin Feng, Shu-Jing Song, Shun-Ai Liu, Hong-Shan Wei

    Jie Yan, Wen Xie, Xin Feng, Shu-Jing Song, Shun-Ai Liu, Hong-Shan Wei, Beijing Ditan Hospital, Beijing 100011, China

    Lei Wang, Jing-Bo Wang, Ji’nan Hospital of Infectious Diseases, Medical College of Shandong University, Ji’nan 250021, Shandong Province, China

    Supported by the Capital Medical Science Foundation, No. 2002-3046

    Correspondence to: Jie Yan, Beijing Ditan Hospital, Beijing 100011, China. jieyan@bbn.cn

    Received: 2006-02-06 Accepted: 2006-02-16

    Abstract

    AIM: To establish a simple, accurate and practical method for the detection of adefovir dipivoxil resistance-associated mutation (rtA181V mutation) in hepatitis B virus.

    METHODS: Four shares of serum were collected from patients who had been treated with adefovir dipivoxil for more than 1 year, and HBV DNA replication reoccurred. Two pairs of primers were designed to amplify the region of HBV reverse transcriptase codon 181(rt181) in order to introduce a BlpI restriction site upon PCR product of wild type (wt). After amplification, the PCR products were digested with BlpI and then subjected to electrophoresis. The patterns of rtA181V mutation were detected by restriction fragment length polymorphism (RFLP) analysis.

    RESULTS: Of the 4 patients, rtA181V mutation was detected in 2 cases by the above method ......

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