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肝癌细胞系(HepG2)感染HCV RNA的研究

THE STUDY OF THE INFECTION OF HEPATITISC VIRUSRNA IN THE HepG2 CELL LINE IN VITRO

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【作者】 叶进杨木兰曾令兰刘微高勇罗端德

【Author】 Ye Jin,Yang Mulan* ,Zeng Linglan,Liu Wei,Gao Yong,Luo Duande ( Departmentof Infectious Disease,Xiehe H ospital,Tongji Medical University,Wuhan 430 0 2 2 , * Departmentof Ultrastructural Pathology,Tongji Medical University,Wuhan 430 0 30 ,China)

【机构】 同济医科大学附属协和医院传染科!武汉430022同济医科大学实验中心超微病理学研究室!武汉430030同济医科大学附属协和医院传染科!武汉4300

【摘要】 为建立丙型肝炎病毒 (HCV )体外感染和细胞培养系统 ,用定量的 HCV RNA阳性血清感染人肝癌细胞系 (Hep G2细胞系 ) ,应用地高辛标记 HCV RNA探针原位杂交技术和 RT- PCR方法对感染后的细胞和上清液中的 HCV RNA进行了检测。在感染后的第一代至第七代的细胞中出现特异性杂交阳性信号 ,第一代、第二代和第六代检测出 HCV RNA正链 ,并在感染后第一、二代检测出 HCV RNA负链。显示 HCV不仅能在体外感染 Hep G2细胞系 ,而且有基因的复制 ,证明 Hep G2细胞能作为 HCV的体外细胞培养系。

【Abstract】 To establish a cell line as a cultural system for HCV infection and propagation in vitro,a human Hep G2 cell line was incubated with a HCV RNA positive serum.The digoxigenin labelled probe by in situ hybridization and polymerase chain reaction amplification were employed to examine the viral RNA in those cells and in its culture medium.The special hybridization positive signals appeared after infection on the Ist subculture to the 7th subculture and viral plus- standed RNA was detected on the 1 st,2 ndand 6th subculture and viral minus- standed RNA was detected on the 1 st and 2 nd subculture post incubation.The cells may have been infected by HCV virus and the virus may have replicated in cell.Therefore,the Hep G2 cell line may be served as a potential host for establishmentof HCV infection in vitro.

【关键词】 原位杂交感染丙型肝炎病毒
【Key words】 In situ hybridizationInfectionHepatitis C virusHuman
【基金】 国家教委留学回国人员基金资助!教外司留 [No.1995(80 6 ) ]
  • 【文献出处】 中国组织化学与细胞化学杂志 ,CHINESE JOURNAL OF HISTOCHEMISTRY AND CYTOCHEMISTRY , 编辑部邮箱 ,2000年03期
  • 【分类号】R392.1
  • 【下载频次】47
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