中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2011年
7期
516-520
,共5页
梁盼盼%郭进军%李青岭%罗强%石小枫%黄爱龙
樑盼盼%郭進軍%李青嶺%囉彊%石小楓%黃愛龍
량반반%곽진군%리청령%라강%석소풍%황애룡
肝炎病毒,乙型%拉米夫定%突变%耐药%病毒准种
肝炎病毒,乙型%拉米伕定%突變%耐藥%病毒準種
간염병독,을형%랍미부정%돌변%내약%병독준충
Hepatitis B virus%Lamivudine%Mutation%Resistance%Virus quasispecies
目的 探讨拉米夫定耐药后换用恩替卡韦补救治疗的慢性乙型肝炎患者的病毒准种演变.方法 提取1例慢性乙型肝炎患者治疗中的7个不同时间点(0、24、48、60、72、96、152周)血清中的HBV DNA,巢式聚合酶链反应法扩增HBV DNA聚合酶基因逆转录酶区,克隆测序法对逆转录酶区氨基酸替换形式及准种分布进行分析,并采用扩增耐药突变系统聚合酶链反应法对患者病毒种群中野毒株与病毒总量进行定量检测.结果 患者在治疗过程中主要存在rtM204V、rtL180M+rtM204V和rtM204I 3种拉米夫定耐药相关的病毒株变异形式,各病毒株所占比例不断发生变化,基线时HBV野毒株为优势病毒株,在病毒学突破时,种群中全部为耐药突变株;换用恩替卡韦治疗后,随着病毒载量的下降,拉米夫定耐药突变株被抑制,野毒株在种群中比例逐渐上升,并成为优势病毒株(79.3%).扩增耐药突变系统聚合酶链反应检测结果显示,在基线和发生病毒学突破时,野毒株在种群中的比例分别为68.55%和0.21%,换药治疗后24周,野毒株比例开始上升,此后野毒株占种群的比例波动于16.01%~26.93%.结论 慢性乙型肝炎患者在核苷(酸)类药物序贯治疗过程中,HBV种群的准种分布一直处于动态变化中.不同的HBV准种演变模式可能在恩替卡韦补救治疗中对恩替卡韦耐药发生的作用也不相同.
目的 探討拉米伕定耐藥後換用恩替卡韋補救治療的慢性乙型肝炎患者的病毒準種縯變.方法 提取1例慢性乙型肝炎患者治療中的7箇不同時間點(0、24、48、60、72、96、152週)血清中的HBV DNA,巢式聚閤酶鏈反應法擴增HBV DNA聚閤酶基因逆轉錄酶區,剋隆測序法對逆轉錄酶區氨基痠替換形式及準種分佈進行分析,併採用擴增耐藥突變繫統聚閤酶鏈反應法對患者病毒種群中野毒株與病毒總量進行定量檢測.結果 患者在治療過程中主要存在rtM204V、rtL180M+rtM204V和rtM204I 3種拉米伕定耐藥相關的病毒株變異形式,各病毒株所佔比例不斷髮生變化,基線時HBV野毒株為優勢病毒株,在病毒學突破時,種群中全部為耐藥突變株;換用恩替卡韋治療後,隨著病毒載量的下降,拉米伕定耐藥突變株被抑製,野毒株在種群中比例逐漸上升,併成為優勢病毒株(79.3%).擴增耐藥突變繫統聚閤酶鏈反應檢測結果顯示,在基線和髮生病毒學突破時,野毒株在種群中的比例分彆為68.55%和0.21%,換藥治療後24週,野毒株比例開始上升,此後野毒株佔種群的比例波動于16.01%~26.93%.結論 慢性乙型肝炎患者在覈苷(痠)類藥物序貫治療過程中,HBV種群的準種分佈一直處于動態變化中.不同的HBV準種縯變模式可能在恩替卡韋補救治療中對恩替卡韋耐藥髮生的作用也不相同.
목적 탐토랍미부정내약후환용은체잡위보구치료적만성을형간염환자적병독준충연변.방법 제취1례만성을형간염환자치료중적7개불동시간점(0、24、48、60、72、96、152주)혈청중적HBV DNA,소식취합매련반응법확증HBV DNA취합매기인역전록매구,극륭측서법대역전록매구안기산체환형식급준충분포진행분석,병채용확증내약돌변계통취합매련반응법대환자병독충군중야독주여병독총량진행정량검측.결과 환자재치료과정중주요존재rtM204V、rtL180M+rtM204V화rtM204I 3충랍미부정내약상관적병독주변이형식,각병독주소점비례불단발생변화,기선시HBV야독주위우세병독주,재병독학돌파시,충군중전부위내약돌변주;환용은체잡위치료후,수착병독재량적하강,랍미부정내약돌변주피억제,야독주재충군중비례축점상승,병성위우세병독주(79.3%).확증내약돌변계통취합매련반응검측결과현시,재기선화발생병독학돌파시,야독주재충군중적비례분별위68.55%화0.21%,환약치료후24주,야독주비례개시상승,차후야독주점충군적비례파동우16.01%~26.93%.결론 만성을형간염환자재핵감(산)류약물서관치료과정중,HBV충군적준충분포일직처우동태변화중.불동적HBV준충연변모식가능재은체잡위보구치료중대은체잡위내약발생적작용야불상동.
Objective To investigate the evolution of hepatitis B virus (HB V) quasispecies in one patient during lamivudine (LAM) monotherapy and switching to entecavir (ETV) rescue treatment. Methods Serum samples were taken at seven different time points during antiviral therapy (0, 24, 48, 60, 72, 96,152 weeks, respectively), the HBV DNA polymerase gene was amplified, cloned and sequenced to analyze the amino acid substitutions within HBV DNA polymerase gene and distribution of virus quasispecies. Quantitative detection of the HBV wild strains and total virus was performed by amplification refractory mutation system real-time PCR (ARMS-PCR). Results Three mutation patterns detected during antiviral therapy in the patient: rtM204V, rtM204V+rtL180M and rtM204I. The HBV quasispecies were found always in dynamic variation. The HBV populations were completely replaced with the LAM-resistant variants when the viral breakthrough was encountered during LAM monotherapy. Interestingly, the wild-type variants presented gradually dominant (79.3%) with the decline of HBV DNA load after switching to ETV rescue administration. ARMS-PCR results showed that the wild-type variants account ed for 68.55% of the HBV populations at baseline and this proportion declined to 0.21% when the viral breakthrough emerged under LAM therapy. The wild-type variants gradually increased from week 24 after switching to ETV rescue therapy and the proportion of HBV wild-type variants in the population fluctuated between 16.01% to 26.93%. Conclusions The distribution of virus quasispecies were always in dynamic variation during sequential therapy with nucleotide analogs in chronic hepatitis B patients. Different patterns of dynamic HBV quasispecies may have different contribution in ETV resistance in LMV refractory patients with ETV administration.