摘要
目的建立具有高感染性的丙型肝炎病毒(HCV)的体外细胞株。方法构建高表达CD81的Huh7细胞株。将HCV质粒(JFH-1,2a)体外转录成RNA,电转染到Huh7和CD81-Huh7细胞中,实时定量PCR法检测两组细胞和培养上清液中病毒载量,间接免疫荧光法检测细胞中HCV核心蛋白表达。收集电转后细胞上清液重新感染两组细胞,检测细胞中病毒RNA表达。结果 CD81-Huh7电转后,细胞和上清液中病毒RNA水平均明显高于Huh71-2log。其被感染的效率也高于Huh7。结论成功建立一株能够高表达感染性HCV病毒颗粒的细胞株CD81-Huh7。
Objective To establish a stable cell line which can enhance HCV infection and virus production.Methods CD81 molecule highly expressed cell line (CD81-Huh7) was established.The HCV RNA (JFH-1,2a) was transcribed in vitro and transfected into Huh7 and CD81-Huh7 by electroporation.HCV RNA was detected by Real time PCR,HCV core protein was observed by immunofluorescence assay.Fresh CD81-Huh7 and Huh7 cells were re-infected by collected supernatants.Results HCV RNA levels in the supernatant and CD81-Huh7 cells were significantly higher than that in the supernatant and Huh7 cells.The supernatants of CD81-Huh7 were infectious and more CD81-Huh7 cells were re-infected than Huh7 cells.Conclusions A CD81 over-expressed cell line which can enhance HCV production in vitro was established in our Lab.
出处
《北京医学》
CAS
2010年第6期424-427,共4页
Beijing Medical Journal
基金
国家"863计划"基金(2007AA02Z151)
北京市优秀人才培养项目(20071B0301600105)