苏南地区1b型丙型肝炎病毒包膜区变异进化研究  

Study On the Variation of E2 of 1b Hepatitis C Virus in the South of Jiangsu Province

作　　者：许可[1] 蒋春梅[1] 邓小昭[2] 丁伟良 谈永飞 张云[1] 

机构地区：[1]南京医科大学流行病与卫生统计学系,江苏南京210029 [2]南京军区军事医学研究所,江苏南京210002 [3]宜兴市第一人民医院,江苏无锡214200

出　　处：《中国病毒学》2006年第4期333-338,共6页Virologica Sinica

基　　金：江苏省高校自然科学研究指导性计划项目(03KJD330144);江苏省自然科学基金项目(BK2004011)

摘　　要：本研究旨在了解本地区丙型肝炎病毒(Hepatitis C virus,HCV)基因型构成的前提下,分析1b型丙型肝炎病毒包膜2(second envelope glycoprotein E2)区的变异和种系进化,并研究其准种变异与临床肝病活动度的关系。对宜兴市人民医院收集的抗HCV抗体阳性患者166名,RT-PCR方法检测HCVRNA,HCVRNA阳性患者采用型特异性引物分型法确定病毒基因型;选择其中未经干扰素治疗的43例1b型慢性丙型肝炎患者的血清标本,扩增E2区,从中选取肝硬化患者4例,慢性非肝硬化患者6例的E2区PCR产物纯化测序,序列采用CLUSTALW与GENBANK上多株不同型别的HCV序列进行比对分析,结果采用Phylip软件构建遗传进化树;并观察E2区高变区1(HVR-1)的氨基酸(aminoacid,aa)残基序列的变异特征;采用单链构象多态性垂直电泳检测43例患者个体内HCVE2区准种的变异情况,比较不同肝病活动度患者准种变异情况。结果表明本地区HCV以1b型为主(84.3%),对E2区基因序列和氨基酸序列变异的分析显示其变异具有一定的规律性,种系进化树提示本地区HCV病毒序列与上海、湖南、日本等地的HCV株有较近的亲缘性。43例患者中ALT高于正常的丙型肝炎患者准种复杂程度明显高于ALT正常者(P<0.05)。故本地区HCV基因变异符合中国东南部的特点,基因变异与临床肝病活动度具有相关性。The aim of this study was to investigate the variation of the second envelope glycoprotein E2 of Hepatitis C Virus strains from the south of Jiangsu province and to learn about the relationship between HCV quasispecies and alanine aminotransferase （ALT）. Identification genotypes of sera samples obtained from 166 donors who were HCV positive was carded our by the PCR with type specific primers based on the sequence of 5′non-coding region （5 ′ NCR）. Forty-three chronic lb type hepatitis C patients who had not received any antiviral therapy were categorized into two groups according to the ALT level. HCV quasispecies diversity were determined by RT-PCR single-strand conformational polymorphism analysis of E2/NS1. Ten strains of HCV E2 were sequenced. A phylogenetic tree of HCV E2 sequences was constructed by using CLUSTAL W and PHYLIP. Out of total 166 patients, 102 were HCV RNA positive in which 86 patients had genotype lb（86/102; 84.3%）, 6 patients had genotype 2 （6/102; 5.9%）, 5 patients waslb/2 mixed types （5/102; 4.9%） and a genotype could not be determined in 5 patients. Phylogeny confirmed that variation in the HCV E2 region was not random but with several conservative gene sequences and amino acid in fixed sites. The quasispecies bands numbers for 43 strains were significantly different with 2.54±1.05 and 4.48± 2.14, respectively. Logistic regression analysis also found the quasispecies diversity was an influencing factor in ALT level. Type lb was the predominant HCV genotype in this area. Phylogenetic trees of HCV E indicated that HCV strains in south of JIANGSU province was close to the ones from SHANGHAI and JAPAN. Some conservative sites were fixed in HCV E region. Quasispecies diversity was correlated with ALT level.

关 键 词：丙型肝炎病毒 包膜区2 进化树 准种 丙氨酸转氨酶 

分 类 号：R373[医药卫生—病原生物学]