恩施地区乙型肝炎患者中乙型肝炎病毒基因组核苷类似物耐药突变研究

Study on nucleotide analogue-resistant mutations in hepatitis B viral genomes in patients with hepatitis B inEnshi

目的 探讨慢性乙型肝炎病毒（HBV）逆转录酶（RT）区的耐药突变与和核苷类似物耐药性的发展之间的相关性.方法 收集恩施自治州中心医院2006~2015年之间使用核苷类似物治疗的乙型肝炎患者.NA治疗的CHB患者样本中编码完整的逆转录酶区（RT的）乙型肝炎病毒基因片段通过聚合酶链反应扩增,PCR产物直接测序并对RT 基因突变进行分析.比较患者中HBV基因型和RT变异.结果 检查出15个NA相关突变位点和6个新的突变.在这些患者中,205例（74.8% ）出现病毒学突破;42例（87.5% ）有部分/无病毒学应答.272例（85% ）携带至少一个 HBV耐药突变.突变组中基因型耐药率显著高于野生型组（P〈0.001）.此外,采用核苷类似物联合治疗的85例（85 /99,85.8% ）实现了部分/无病毒学应答.联合治疗与无基因型耐药与较高的病毒学应答率相关.结论 这些数据表明耐药突变可能是经历病毒学突破接受核苷类似物患者治疗失败的主要原因.一旦有迹象显示初始抗病毒治疗无效,临床医生应及时进行基因型耐药测试.

Objective To explore the correlation between chronic hepatitis B virus （HBV） drug-resistant mutation in reverse transcriptase （RT） region and nucleoside analog resistance development in patients with antiviral treatment.Methods Patients with nucleoside analog treatment were recruited between 2006 and 2015 from Central Hospital of Enshi Autonomous Prefecture hospital,Enshi,People′s Republic of China.HBV gene fragment encompassing the complete reverse transcriptase （RT）gene in NA-treated CHB patient samples was amplified by polymerase chain reaction and PCR product-based direct sequencing was performed. Mutations in the RT gene were analyzed.Patients were identified with HBV-B and HBV-C genotype infections,HBV genotype and RT mutation were compared.Results 15 reported NA related mutation sites and 6 novel mutations were detected.Among the pa-tients,205 patients （74.8% ）experienced virological breakthrough;42 patients （87.5% ）had partial/null response.272 （85% ）car-ried at least one HBV drug resistance mutation.The prevalence of genotypic resistance was significantly higher in mutation group than in wild-type group （P〈0.001）.In addition,85 patients （85/99,85.8% ）achieved had partial/null response with the nucleo-side analog combination therapy.Combination therapy and no evidence of genotypic resistance were related to a higher rate of viro-logical response.Conclusion These data indicate demonstrates that drug-resistant mutations may be the main cause of nucleoside analog treatment failure in patients experiencing virological breakthrough.Once patients treated with initial antiviral treatment show signs of inefficacy,clinicians should perform genotypic resistance testing early.