广东HCV进化与注射吸毒者HCV NS5B区正向选择位点的分析研究  

HCV evolution and positive selection sites of HCV NS5B region among intravenous drug users in Guangdong

作　　者：王敏[1] 许茹[1] 廖峭[1] 钟惠珊 黄杰庭[1] 杜荣松[1] 单振刚[1] 戎霞[1] 付涌水[1] WANG Min;XU Ru;LIAO Qiao;ZHONG Huishan;HUANG Jieting;DU Rongsong;SHAN Zhengang;RONG Xia;FU Yongshui(Institute of Blood Transfusion,Guangzhou Blood Center,the Key Medical Laboratory of Guangzhou(2021-2023),Guangzhou 510095,China)

机构地区：[1]广州血液中心广州市血液安全重点实验室,广东广州510095

出　　处：《中国输血杂志》2022年第6期597-601,共5页Chinese Journal of Blood Transfusion

基　　金：国家自然科学基金(32000666);广东省自然科学基金(2020A1515010118);广州市医学重点学科项目(2021~2023)。

摘　　要：目的了解广东HCV进化与注射吸毒者(IDUs)HCV NS5B区正向位点的选择压力情况。方法随机收集广东141(人)份HCV RNA阳性献血者和58(人)份HCV患者标本做HCV NS5B序列扩增,结合前期测序得到的2009年~2011年的献血者和IDUs HCV NS5B序列,用分子进化遗传学分析(MEGA)软件做同源性分析,贝叶斯进化分析抽样树(BEAST)软件包做进化分析,用Datamonkey在线软件包进化的混合效应模型(MEME)方法对IDUs分离的序列做选择压力分析,用进化分析软件Arlequin的Tajima和Fu中性检验分析种群扩张情况。结果本组IDUs不同亚型内部同源性比较结果为:HCV-3b同源性最高(97%),其次为HCV-3a(96%),HCV-6a(95%)和HCV-1b(94%);HCV进化速率分析显示:HCV-1b的进化速率最快[2.17E-03替换数/位点/年(y/y/y)],其次为HCV-3b(2.12E-03 y/y/y),HCV-3a(1.58E-03 y/y/y)和HCV-6a(1.28E-03 y/y/y);HCV理论感染人数分析:1980年~1990年是HCV-6a快速增长期,1990年~1995年是HCV-1b快速增长期,2000年~2007年是HCV-3a快速增长期;HCV群体遗传特征:HCV-1b、3a、3b和6a均经历了种群扩张,其中3a和3b最明显。HCV选择压力分析:IDUs NS5B序列中339个核苷酸有2个正向选择位点(第235和243位点),但此段基因的5个直接抗病毒药物(DAA)位点仅在第316位点发生了突变[突变率1.24%(14/1130)]。结论广东地区HCV 3b进化呈明显的种群扩张趋势,尤其在当地IDUs中的比例、同源性较高,应作为本地区HCV防治的重点。鉴于广东IDUs HCV NS5B基因片段的正向选择位点均非DAA结合位点,因此设想DAA对这些患者有较好的疗效。Objective To learn the situation of the evolution process of HCV virus population and the selection pressure of HCV NS5 B in intravenous drug users(IDUs)in Guangdong.Methods 141 blood samples from hepatitis C virus(HCV)RNA-positive blood donors and 58 from HCV patients in Guangdong were randomly collected for HCV NS5B sequence amplification,combined with HCV NS5B sequences from blood donors and IDUs obtained by sequencing previously(between 2009 and 2011).Homology analysis was performed by Molecular Evolutionary Genetics Analysis(MEGA)software,evolutionary analysis were performed by Bayesian Evolutionary Analysis Sampling Trees(BEAST)software package.Selection pressure analysis was performed on sequences isolated from IDUs by Datamonkey online software package with Mixed Effects Model Evolution(MEME)method,and the population expansion of species were analyzed using Tajima and Fu neutrality test by Arlequin software.Results The comparison results of internal homology among different subtypes of IDUs in this group were as follows:HCV-3b had the highest homology(97%),followed by HCV-3a(96%),HCV-6a(95%)and HCV-1b(94%);HCV evolution rate analysis showed that HCV-1b had the fastest evolution rate[2.17 E-03 substitutions/site/year(y/y/y)],followed by HCV-3b(2.12 E-0 y/y/y),HCV-3a(1.58 E-03 y/y/y)and HCV-6a(1.28 E-03 y/y/y).The analysis on effective population of HCV:1980~1990 was rapid growth period for HCV-6a,1990~1995 period for HCV-1b,and 2000~2007 period for HCV-3a.HCV population genetic characteristics was as follows:HCV-1b,3a,3b and 6a experienced population expansion,among which 3a and 3b were the most obvious.As to the analysis of HCV selection pressure,two positive selection sites(235 and 243)were found in the 339 nucleotide fragment of the NS5B sequence in injecting drug users,but mutation only occurred at position 316[mutation rate 1.24%(14/1130)]among 5 direct antiviral drug(DAA)sites in this gene.Conclusion The evolution of HCV-3b in Guangdong has showed an obvious trend of population expansion,with a high

关 键 词：丙型肝炎病毒 HCV进化 HCV选择压力 注射吸毒者 正向选择位点 直接抗病毒药物 广东 

分 类 号：R446[医药卫生—诊断学] R512.63[医药卫生—临床医学]