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目的探讨丙型肝炎病毒(HCV)体外感染树鼩原代肝细胞。方法用HCV RNA阳性血清感染原代树鼩肝细胞,并用感染后细胞培养上清液进行传代感染,通过检测受染肝细胞正、负链HCV RNA、培养上清液中包装后HCV RNA,并对比分析感染前后病毒准种变化等指标,评价感染是否成功。结果受染树鼩肝细胞自第5-10天可检出负链HCV RNA,而正链RNA至感染后第14天仍可检出;感染后3-14 d培养上清液中可检出HCV RNA,且呈RNA酶抗性;培养上清液中病毒可传代感染新的树鼩肝细胞;感染前后HCV准种分析显示树鼩肝细胞可被特定的准种选择性感染,而传代感染后肝细胞可检出新的准种。结论原代树鼩肝细胞体外可被HCV感染且支持病毒复制。
Objective To investigate hepatitis C virus (HCV) in vitro infection of tree shrew primary hepatocytes. Methods Primary hepatocytes were infected with HCV RNA positive sera and subcultured with supernatants of infected cells. The positive and negative HCV RNA of infected hepatocytes were assayed, and the packaged HCV RNA in supernatants Comparative analysis of indicators of changes in the virus quasispecies before and after infection, evaluation of infection is successful. Results The infected shrew hepatocytes detected negative-sense HCV RNA from day 5-10, while the positive-strand RNA was still detectable on the 14th day after infection. After 3-14 days of infection, the supernatant could be detected HCV RNA, and showed RNase resistance; the virus in the culture supernatant could be passaged to infect new tree shrew hepatocytes; quasispecies analysis of HCV before and after infection showed that tree shrew hepatocytes could be specifically quasi-selectively infected while passage infection After the liver cells can detect new quasi-species. Conclusion The primary tree shrews can be infected with HCV in vitro and support virus replication.