Paulson Matthew S
Gilead Sciences, Foster City, California, USA.
Curr Protoc Pharmacol. 2010 Dec;Chapter 13:Unit 13B.4. doi: 10.1002/0471141755.ph13b04s51.
The development of robust genotype 1b and genotype 1a hepatitis C virus (HCV) replicon systems has enabled the convenient in vitro study of part of the virus life cycle. This unit describes detailed protocols for generating and measuring infectious HCV, or cell-culture-derived infectious HCV (HCVcc). The HCVcc infectious system has two essential components: (1) cells that are permissive to de novo infection and allow effective replication of the full virus life cycle; and (2) a virus genome that has robust and efficient replication in tissue culture. The assays in this unit are based on protocols designed for Huh-7-derived cell lines that allow robust replication of HCV and are permissive to infection. These protocols are important for the implementation of drug discovery efforts relative to the entire infectious virus life cycle.
强大的1b型和1a型丙型肝炎病毒（HCV）复制子系统的开发，使得对病毒生命周期部分环节进行便捷的体外研究成为可能。本单元描述了生成和检测感染性HCV或细胞培养衍生感染性HCV（HCVcc）的详细方案。HCVcc感染系统有两个基本组成部分：（1）允许从头感染并能使完整病毒生命周期有效复制的细胞；（2）在组织培养中具有强大而高效复制能力的病毒基因组。本单元中的检测方法基于为源自Huh-7的细胞系设计的方案，这些细胞系能使HCV进行强大的复制并允许感染。这些方案对于开展与整个感染性病毒生命周期相关的药物研发工作至关重要。
