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乙肝病毒多聚酶区rtL229突变的演变及其表型分析
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乙肝病毒多聚酶区rtL229突变的演变及其表型分析
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Journal Article
乙肝病毒多聚酶区rtL229突变的演变及其表型分析
2012
Overview
目的分析乙肝病毒（HBV）多聚酶的反转录酶（RT）功能域rtL229突变的演变规律及其与拉米夫定（LAM）耐药的相关性。方法回顾性分析2010年8月在解放军302医院抗病毒治疗过程中发生rtL229F突变的1例慢性乙肝患者的临床资料,从其动态血清中扩增HBV RT基因并克隆测序（〉20个克隆/样本）以观测rtL229F突变的演变过程。同时将含有不同突变形式（rtM204I、rtL229F以及rtM204I＋rtL229F）的HBV DNA RT区基因定向克隆至HBV1.1载体中获得可复制的HBV复制子,将复制子转染HepG2细胞,在含或不含核苷（酸）类似物[LAM、阿德福韦酯（ADV）、恩替卡韦（ETV）和替诺福韦酯（TDF）]的培养基中连续培养4d,采用实时荧光定量PCR法检测上清中产生的HBV DNA。结果 LAM治疗过程中,rtL229F突变可继发于rtM204I突变,而停用LAM后可逐渐回复为野生型。表型分析结果显示,单独rtL229F突变不改变病毒的药物敏感性,而与rtM204I突变联合后可以增强突变HBV的LAM耐药性。结论 rtL229F位点突变是LAM耐药突变的补偿突变,与LAM应答不佳相关,且对ADV、ETV及TDF敏感。
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