摘要
核苷(酸)类似物是临床上治疗慢性乙型肝炎病毒(HBV)感染最常用的药物,但长期应用可引发病毒耐药,导致治疗失败。基因变异检测及表型耐药分析是发现和鉴定HBV耐药的基本方法,前者主要是应用基因测序或线性反向探针杂交等方法,检出已知的病毒耐药相关突变,后者则是在体外细胞水平确定携带有变异基因的HBV毒株对病毒复制力及核苷(酸)类药物敏感性的影响,是鉴定复杂与特殊HBV耐药变异的基本手段。我们通过技术创新,建立了敏感、特异、经济、高效的HBV基因型检测方法,进而建立了可靠的HBV复制力和表型耐药分析方法,广泛应用于临床样本分析,实现了对HBV耐药的早期发现及对新的非典型HBV耐药相关突变株的鉴定。我们近期在核苷(酸)类似物耐药和应答不佳的患者中检测到多种特殊和复杂HBV耐药相关突变,并结合临床资料,进一步对代表性变异株进行病毒复制力和表型耐药分析,取得了多项重要发现:①从大样本中检出和鉴定了多种多重耐药HBV变异株,并发现联合用药可协同抑制多重耐药HBV变异株的体外复制;②HBV rtL229替换可作为补偿变异,恢复拉米夫定耐药变异株rtM204I的病毒复制力;③rtM204Q是一种新的拉米夫定耐药相关突变;④rtA181C+rtL180M+rtM204V与恩替卡韦耐药相关;⑤同一耐药患者血清HBV虽检出耐药变异,但外周血单个核淋巴细胞(PBMCs)中的HBV cccDNA仍以原始野生株为优势种群,提示PBMCs为体内HBV野生株的"存储库",参与HBV肝外感染。上述发现对深入揭示HBV耐药变异的临床特点和发生机制,辅助临床合理制定并优化抗病毒治疗方案具有重要作用。
Nucleoside and nucleotide analogs are most commonly-used agents in the treatment of hepatitis B virus（HBV） infection.However,long-term use of nucleoside/nucleotide analogs may induce HBV drug resistance,thereby leading to treatment failure.The genotyping and phenotyping are two basic methods for detection and identification of HBV drug resistance.Genotyping is used to detect well-known drug-resistant mutations mainly through DNA sequencing and reverse hybridization line probe.Phenotyping is an essential tool to identify ＂novel＂ and complex resistant mutations,based on comparison of HBV replication capacity and drug susceptibility between variants and wild-type counterparts.We have developed sensitive,specific,high costeffectiveness assay for HBV genotypic resistance detection,thus developing reliable assays for analyses of HBV replication capacity and phenotypic resistance.These assays have been widely-applied in clinic,realizing early finding of classical drug-resistant mutations and identification of unusual drug-resistant mutations.In our recent studies,we detected some unusual and complex HBV mutations in the patients with resistance and without good response to nucleos（t）ide analogs.In combination with clinical information and phenotyping of replication capacity and drug susceptibility,we have acquired novel findings as follows.（1） Detected and identified several multidrug-resistant HBV strains by screening of a large number of patients;and multidrug-resistant HBV strains could be synergistically suppressed by combined use of anti-HBV agents.（2） It was found that HBV rt229 substitutions could be a compensatory mutation to increase replication capacity of lamivudine-resistant rtM204I mutant.（3） rtM204Q was found as a novel lamivudine-resistance-associated mutation.（4） rtA181C＋rtL180M＋rtM204V was found to be associated with entecavir resistance.（5） It was found that in the same patient with drug-resistant HBV strains in sera,wild-type HBV strains were predominant in cccDNA of their peripheral blood mononuclear cells（PBMCs）,suggesting that PBMCs might be a storage pool of wild-type HBV associated with extra-hepatic HBV infection mechanism.These findings are important for a better understanding of clinical features and underlying mechanism of HBV drug-resistant mutations,capacitating clinicians to optimize antiviral schedule.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2012年第6期535-538,共4页
Medical Journal of Chinese People's Liberation Army
基金
国家"十二五"传染病重大专项子课题(2012ZX10004503)
国家自然科学基金面上课题(81171617)
军队医院"十二五"临床高新技术项目(2010gxjs009)~~
关键词
肝炎病毒
乙型
突变
抗药性
病毒
表型
hepatitis B virus
mutation
drug resistance
viral
phenotype
作者简介
徐东平．医学博士，研究员，博士生导师、全军传染病研究所病毒性肝炎研究室（军队重点实验室）主任，解放军302医院肝衰竭诊疗与研究中心病毒性肝炎研究室主任．中国免疫学会感染免疫分会委员．1999—2002年美国纽约大学西奈山医学院访问学者，现主要研究方向为病毒性肝炎的发病机制、病毒耐药与免疫治疗．近年承担国家传染病重大专项、国家973、国家自然科学基金、北京市自然科学基金重点课题及军队杰出人才课题等共16项；发表论文180余篇．其中以主要作者在New England Journal of Medicine, Gastroenterology, Journal of Immunology, Journal of ViralHepatitis, Journal of Clinical Virology, Journal of Clinical Microbiology等SCI收录期刊上发表论文45篇获国家科技进步二等奖2项，中华医学会科学技术一等奖1项，军队医疗成果一等奖1项．军队或北京市科研成果二等奖4项。
