拉米夫定治疗慢性乙型肝炎中变异病毒株的动态变化
乙型肝炎病毒;变异;拉米夫定;焦磷酸测序,田国保,席宏丽,田地,崔建军,于敏,陆海英,曾争,通讯作者:,DynamicsofhepatitisBvirusvariationsduringlamivudinetreatment,Guo-BaoTi
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田国保, 席宏丽, 田地, 崔建军, 于敏, 北京大学第一医院感染疾病科病毒研究室 北京市 100034
陆海英, 曾争, 北京大学第一医院感染疾病科 北京市 100034
通讯作者: 曾争, 100034, 北京市, 北京大学第一医院感染疾病科. zeng@bjmu.edu.cn
电话: 010-66551799
收稿日期: 2007-04-15 修回日期: 2007-08-18
Dynamics of hepatitis B virus variations during lamivudine treatment
Guo-Bao Tian, Hong-Li Xi, Di Tian, Jian-Jun Cui, Min Yu, Hai-Ying Lu, Zheng Zeng
Guo-Bao Tian, Hong-Li Xi, Di Tian, Jian-Jun Cui, Min Yu, Laboratory of Virology, Department of Infectious Diseases, Peking University First Hospital, Beijing 100034, China
Hai-ying Lu, Zheng Zeng, Department of Infectious Disease, Peking University First Hospital, Beijing 100034, China
Correspondence to: Zheng Zeng, Laboratory of Virology, Peking University First Hospital, Beijing 100034, China. zeng@bjmu.edu.cn
Received: 2007-04-15 Revised: 2007-08-18
Abstract
AIM: To use pyrosequencing to establish a method for monitoring hepatitis B virus mutants resistant to lamivudine.
METHODS: Plasmids cloned, sequenced and constructed from the lamivudine-resistant HBV mutants at rtM204I/V (YMDD motif) and rtL180M were used as a standard. The frequencies of mutation-site nucleotides were detected by pyrosequencing in sera from 4 patients who underwent lamivudine therapy. HBV DNA with rtM204I, rtM204V and wildtype plasmids were constructed for subtracting the background signal frequencies.
RESULTS: The frequency of background signals in all plasmids were between 0.1% and 3.9%, The frequency of background signals was determined as 5%. In all 4 patients, HBV variations were detected from 4.5% to 33% before lamivudine therapy. Virological breakthrough was monitored for 4 to 8 months along with the gradually increasing HBV variations at positions of rtM204I/V and rtL180M.
CONCLUSION: Pyrosequencing is an effective method to quantify and monitor HBV variations during lamivudine therapy ......
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