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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 an in vitro cell line with highly infectious hepatitis C virus (HCV). Methods Construction of Huh7 cell line highly expressing CD81. The HCV plasmids (JFH-1,2a) were transcribed into RNA in vitro and electrotransfected into Huh7 and CD81-Huh7 cells. The real-time quantitative PCR was used to detect the viral load in both cell culture supernatants and indirect immunofluorescence assay HCV core protein expression in cells. After electrotransformation, the cell supernatant was re-infected with two groups of cells to detect the expression of viral RNA in the cells. Results After transduction of CD81-Huh7, the levels of viral RNA in the cells and supernatant were significantly higher than that of Huh71-2log. It is also more effective than Huh7. Conclusion A CD81-Huh7 cell line that can express HCV particles with high virulence was successfully established.