INTRODUCTION: Infection with hepatitis C virus is a serious worldwide health problem. Since its discovery in 1989, the development of a cell culture system for HCV has been a major goal for scientists worldwide. In 2005 the first tissue culture that led to the production of HCV particles (2a genotype) has been created. The aim of the study was to determine viral RNA level in an infectious HCV cell culture system. METHODS: Huh-7.5 cell line was infected with the JFH1 (Japanese Fulminant Hepatitis) RNA by lipofection. The level of HCV RNA was measured in supernatant of the cell culture by Real-Time PCR method. RESULTS: HCV RNA was detected in the supernatant of Huh-7.5 cell line infected with the use ofJFH 1 RNA and lipofectamin. The maximal level of HCV RNA (3.69 x 10(9) copies/ml) was detected 96h after transfection. After about 30 days oftransfection, HCV RNA was on the stable level (10(7)-10(8)). CONCLUSIONS: Huh-7.5 cell line infected with HCV form a robust cell culture model of HCV infection. HCV replicates on high levels in Huh-7.5, which is the crucial event for the investigation of new antivirals in in vitro model.
INTRODUCTION: Infection with hepatitis C virus is a serious worldwide health problem. Since its discovery in 1989, the development of a cell culture system for HCV has been a major goal for scientists worldwide. In 2005 the first tissue culture that led to the production of HCV particles (2a genotype) has been created. The aim of the study was to determine viral RNA level in an infectious HCV cell culture system. METHODS:Huh-7.5 cell line was infected with the JFH1 (Japanese Fulminant Hepatitis) RNA by lipofection. The level of HCV RNA was measured in supernatant of the cell culture by Real-Time PCR method. RESULTS: HCV RNA was detected in the supernatant of Huh-7.5 cell line infected with the use ofJFH 1 RNA and lipofectamin. The maximal level of HCV RNA (3.69 x 10(9) copies/ml) was detected 96h after transfection. After about 30 days oftransfection, HCV RNA was on the stable level (10(7)-10(8)). CONCLUSIONS:Huh-7.5 cell line infected with HCV form a robust cell culture model of HCV infection. HCV replicates on high levels in Huh-7.5, which is the crucial event for the investigation of new antivirals in in vitro model.