解放军医学杂志2012，Vol.37Issue(6)：544-547,4.
乙肝病毒多聚酶区rtL229突变的演变及其表型分析
Evolution and phenotypic analysis of rtL229 polymerase mutations within the hepatitis B virus genome
摘要
Abstract
Objective The rtL229 mutations in the reverse-transcriptase (RT) domain of hepatitis B virus (HBV) polymerase have been reported to have the ability to enhance the replication capacity of lamivudine (LAM)-resistant strains. This study aimed to investigate the evolution and phenotypic characteristics of rtL229 mutations. Methods The clinical characteristics of one representative patient with chronic hepatitis B who had received LAM treatment were retrospectively analyzed. HBV RT genes isolated from different serum samples were amplified by nested PCR, and clonal sequencing (>20 clones/sample) was done to analyze the evolution of rtL229F mutations. Meanwhile, the amplified HBV RT genes bearing different mutation patterns (wild-type, rtM204I, and rtL229F+rtM204I, respectively) were cloned into 1.1-fold HBV vector (pTriEx-mod-l.l) to generate replication-competent viral constructs. The recombinant constructs were transfected into HepG2 cells, which were cultured in the presence or absence of nucleos(t)ide analogs. Supernatant HBV DNA products were quantitated using real-time PCR. Results rtL229F mutation was secondary to rtM204I mutation during LAM treatment, and it regressed to wild-type strains together with rtM204I mutation after LAM withdrawal. Phenotypic assay showed that the single rtL229F mutation had no impact on the drug-susceptibility of HBV to nucleos(t)ide analogs, but could enhance the drug resistance of LAM-resistant rtM204I strain. Conclusion The rtL229F site mutation might be a compensatory mutation of LAM resistance, which is associated with suboptimal response to LAM treatment, and it still is susceptible to the treatment of adefovir dipivoxil, entecavir and tenofovir in vitro.关键词
肝炎病毒,乙型/抗药性,病毒/突变/抗病毒药Key words
hepatitis B virus/ drug resistance, viral/ mutation/ antiviral agents分类
医药卫生引用本文复制引用
纪冬,刘妍,许智慧,李乐,陈丽,李晓东,陈国凤,辛绍杰,徐东平..乙肝病毒多聚酶区rtL229突变的演变及其表型分析[J].解放军医学杂志,2012,37(6):544-547,4.基金项目
国家“十二五”传染病重大专项子课题(2012ZX10004503) （2012ZX10004503）
