甘肃省某市哨点监测人群HCV基因分型及NS5B聚合酶抑制剂耐药相关替代突变分析  

Analysis of HCV genotypes and resistance associated substitutions to NS5B polymerase inhibitors among the sentinel surveillance population in a city from Gansu Province

作　　者：张翠 乔瑞娟[2] 李健[1] 屠爱霞[2] 韩孟杰[1] 王晓蕊[2] 邢文革[1] 梁雯丽 石林[2] 邱茂锋[1] ZHANG Cui;QIAO Ruijuan;LI Jian;TU Aixia;HAN Mengjie;WANG Xiaorui;XING Wenge;LIANG Wenli;SHI Lin;QIU Maofeng(I.National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases,National Center for AIDS/STD Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;Department of AIDS Control and Prevention,Gansu Provincial Center for Disease Control and Prevention,Lanzhou 730020,Gansu,China)

机构地区：[1]中国疾病预防控制中心性病艾滋病预防控制中心传染病溯源预警与智能决策全国重点实验室,北京102206 [2]甘肃省疾病预防控制中心艾滋病防治所,兰州730020

出　　处：《中国艾滋病性病》2024年第9期894-899,共6页Chinese Journal of Aids & STD

基　　金：丙肝防治技术指导与能力建设项目;HIV/HCV分子溯源检测技术及HCV变异株监测预警重点项目。

摘　　要：目的探索建立HCV NS5B区的耐药相关替代突变(RASs)方法并初步应用于甘肃省某市丙肝哨点监测人群。方法以甘肃省某市2020年在丙型肝炎哨点监测中发现的HCV抗体阳性者为研究对象,提取血浆样本中的RNA,经反转录和巢式PCR扩增后进行Sanger测序,分别进行基因亚型和NS5B区耐药相关替代突变分析。结果74份样本中有20份核酸扩增成功并完成测序。所获的HCV基因亚型共有4种,构成比由高到低依次为3a(12/20,60%)、1b(4/20,20%)、2a(3/20,15%)和3b(1/20,5%)。1b亚型的RASs为C316N(4/4)、M414L(1/4)、L392I(1/4)、I424V(1/4)和V499A(1/4),2a亚型的RASs为A421V(3/3),3a亚型的RASs为A150V(9/12),3b亚型未检出RASs。发现耐药的直接抗病毒药物有索磷布韦、达拉他韦和贝拉布韦。结论甘肃省某市哨点监测发现HCV抗体阳性者基因亚型构成呈多样性,普遍存在NS5B聚合酶抑制剂的RASs;不同亚型NS5B区发生的RASs不同,以1b亚型RASs种类最多。Objective To explore the establishment of a resistance associated substitutions(RASs)detection method for hepatitis C virus(HCV)NS5B region,and its preliminary application among a hepatitis C sentinel surveillance population in a city of Gansu Province.Methods Individuals who tested positive for HCV antibodies during the 2020hepatitis C sentinel surveillance in a city in Gansu Province were included in the study.RNA was extracted from plasma samples,and after reverse transcription and nested PCR amplification,Sanger sequencing was performed for HCV subtyping and analysis of RASs in the NS5B region.Results Out of 74 samples,20 were successfully amplified and sequenced.Four HCV genotypes were identified,with a composition ratio,descending from highest to lowest,of 3a(12/20,60%),1b(4/20,20%),2a(3/20,15%),and 3b(1/20,5%).The RASs for genotype 1b included C316 N(4/4),M414L(1/4),L392I(1/4),I424V(1/4),and V499A(1/4);for genotype 2a,A421V(3/3);and for genotype 3a,A150V(9/12).No RASs were detected for genotype 3b.Direct-acting antiviral agents that have been found to be associated with RASs in this study included sofosbuvir,daclatasvir,and beclabuvir.Conclusions Sentinel surveillance in a city in Gansu Province found that HCV antibody-positive individuals have diverse HCV subtypes,with the common presence of RASs for NS5B polymerase inhibitors.The types of RASs that occur in different NS5B subtypes are different,with the 1b subtype having the greatest diversity of RASs.

关 键 词：丙型肝炎病毒 直接抗病毒药物 NS5B聚合酶抑制剂 耐药相关替代突变 Sanger测序 

分 类 号：R373.9[医药卫生—病原生物学] R512.91[医药卫生—基础医学]