CAJ | 학술논문

目的本研究旨在分析乙型肝炎病毒(HBV)阿德福韦(ADV)非经典耐药相关突变的临床流行特点和表型耐药特征.方法选取1741例慢性HBV感染者,PCR直序法检测7种非经典ADV耐药突变.对rtN238T突变患者进行动态克隆测序;构建rtN238T、rtA181V和rtA181V+ rtN238T突变株的pTriEx-HBV 1.1重组载体,瞬时转染HepG2细胞,评价各病毒株的表型耐药特性.结果 rtN238T突变在应用ADV治疗患者中的检出率高于未治疗患者和其他核苷类药物治疗患者(X2=17.10,P＜0.01).随访患者在接受ADV治疗47个月后发生病毒学反弹和生化学突破,并检出rtN238T+ A181V和rtN238T克隆株;随后应用ETV治疗33个月,病毒载量和ALT恢复正常,患者体内病毒全部转变为野生株.rtN238T+ A181V病毒株复制力显著高于rtA181V(t=9.54,P＜0.01).与野生株相比,rtN238T+ A181V突变株对ADV敏感性下降.结论 rtN238T突变可以增加ADV经典耐药突变株rtA181V的复制力,是一种ADV耐药相关的复制力补偿突变.
목적 본연구지재분석을형간염병독(HBV)아덕복위(ADV)비경전내약상관돌변적림상류행특점화표형내약특정.방법 선취1741례만성HBV감염자,PCR직서법검측7충비경전ADV내약돌변.대rtN238T돌변환자진행동태극륭측서;구건rtN238T、rtA181V화rtA181V+ rtN238T돌변주적pTriEx-HBV 1.1중조재체,순시전염HepG2세포,평개각병독주적표형내약특성.결과 rtN238T돌변재응용ADV치료환자중적검출솔고우미치료환자화기타핵감류약물치료환자(X2=17.10,P＜0.01).수방환자재접수ADV치료47개월후발생병독학반탄화생화학돌파,병검출rtN238T+ A181V화rtN238T극륭주;수후응용ETV치료33개월,병독재량화ALT회복정상,환자체내병독전부전변위야생주.rtN238T+ A181V병독주복제력현저고우rtA181V(t=9.54,P＜0.01).여야생주상비,rtN238T+ A181V돌변주대ADV민감성하강.결론 rtN238T돌변가이증가ADV경전내약돌변주rtA181V적복제력,시일충ADV내약상관적복제력보상돌변.
Objective To identify clinical prevalence of untypical adefovir-resistant mutations of hepatitis B virus (HBV),and to analyze their phenotypic characteristics.Methods 1741 patients with chronic HBV infection were evolved.Untypical adefovir-resistant mutations were analyzed by direct sequencing.Longitudinal analysis was performed by clonal sequencing.Wild-type and mutant HBV genomic amplicons were constructed into pTriEx-HBV 1.1 vector and transfected into HepG2 cells.The replication capacity and the 50％ effective concentration of drugs (EC50) were calculated.Results Patients treated with adefovir alone were more likely to develop rtN238T mutation than those treated with other nucleos(t) ide drugs (x2 =17.10,P ＜ 0.01).The patient received adefovir for 47 months,and then viral rebound and biochemical breakthrough occurred with detection of rtN238T + A181V and rtN238T mutation.Switching-to entecavir therapy suppressed HBV DNA and ALT to an undetectable level and converted all viruses into wild type ones.The reulsts of viral replication capacity showed that rtN238T + A181V strain was higher than rtA181V strain (t =9.54,P ＜ 0.01).Compared to the wild type virus,rtN238T + A181V variant was relatively less susceptible to adefovir.Conclusions rtN238T mutation conferred no resistance to ADV but enhanced natural replication capacity,hence it might represent a novel compensatory drug-resistant mutation for adefovir.