慢性肝炎患者血清中丙型肝炎病毒核心区和包膜1区基因变异研究  

Genetic variation of the core and E1 region of hepatitis C virus in serum collected from patients with chronic hepatitis

作　　者：陈嵩[1] 刘志[1] 王宇明[1] 

机构地区：[1]第三军医大学西南医院感染病科,重庆400038

出　　处：《中华肝脏病杂志》2001年第4期220-222,共3页Chinese Journal of Hepatology

基　　金：国家自然科学基金(39870694)

摘　　要：目的探讨丙型肝炎病毒（ＨＣＶ）核心（Ｃ）区和包膜１（Ｅ１）区变异与其感染慢性化的关系。方法１０例ＨＣＶ慢性感染者和２例急性感染者血标本，采用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）扩增ＨＣＶ的Ｃ区羧基端、 Ｅ１及 Ｅ２区氨基端片断（１ｋｂ），扩增产物进行克隆，以单链构象多态性（ＳＳＣＰ）和异质性双体（ＨＤ）分析对每份血清的３０个克隆的Ｃ／Ｅ１区准种（ｑｕａｓｉｓｐｅｃｉｅｓ）进行筛选，挑选每例标本ＨＣＶ的优势株与劣势株序列进行测定，并分析推导的氨基酸序列。结果各例患者血清中ＨＣＶ的Ｃ／Ｅ１区扩增片段形成的ＳＳＣＰ条带间差异明显，而ＨＤ与同源双体之间差距不明显。ＨＣＶ急性感染者和慢性感染者血清病毒Ｃ区无发生变异，后者Ｅ１区氨基酸替换率为１．３２％，但功能性氨基酸无改变。结论ＨＣＶ的Ｅ１区序列变异以准种形式存在，但与免疫逃避可能无关。Objective To study the association of the core and E1 region variation of hepatitis C virus (HCV) with the chronicity of HCV infection. Methods Acute-phase plasma samples from two drug-injection users who acquired HCV infection during six-month follow-up and ten patients with chronic hepatitis C were obtained. A 1-kb fragment containing the 3' half of core, completed E2 and 3' half of E2 were amplified by nested reverse transcription polymerase chain reaction (RT-PCR). For each, 30 cloned cDNAs were examined by a method that combined heteroduplex(HD) analysis and a singlestranded conformational polymorphism(SSCP) assay to assess quasispecies complexity and optimize selection of clones with unique gel shift patterns for sequenceing. Amino acid sequences were analyzed for signature patterns and glycosylation signals. Results There was obvious difference between the SSCP bands of cloned cDNAs representing core-E1 region, but not obvious distance between heteroduplexes and the homoduplex. There was no change in the sequence of each clonotype from acutely HCV-infected subjects, whereas the fate of nonsynonymous substitution was 1 .32% in amino acid sequence of E1 from persistently HCV-infected subjects with except for functional patterns. Conclusions The variation of the E1 gene of HCV is in the presence of quasispecies population, but might not related with its immunologic escape.

关 键 词：丙型肝炎病毒 核心区 包膜区 基因变异 慢性肝炎 

分 类 号：R512.63[医药卫生—内科学]