| Background and objective: Hepatitis C virus is the cause of the hepatitis C, which is a global health problem. According to the World Health Organization Report, More than 180 million people worldwide are infected with hepatitis C virus (HCV), of which 20-50% may develope cirrhosis and 1-2% may cause hepatocellular carcinoma in ten or twenty years. Currently, there is limited effects of combination therapy with interferon-αand ribavirin. Less than 40% of those given this treatment have a durable benefit, and most patients continue to have viremia after the treatment has been stopped. But different genotype achieve different sustained virologic response. So here is an urgent need for more effective therapeutics. However their development has been significantly hamperde by the lack of appropriate cell culture models or small animal model permissive for HCV that allow the propagation of the virus in the laboratory. Recently, the development of selectable dicistronic subgenomic replicons of HCV has provided a new research tool for hepatitis C. These in vitro systems can support efficient HCV RNA replication and synthesis of all viral proteins. Unfortunately complete virus particle has not been achieved yet. HCV RNA genome is approximately 9.6 kb with complicated secondary structure and plenty of G or C base in long viral RNA templates which make HCV long reverse transcriptase polymerase chain reaction (RT-PCR) difficult. Forthermore, technical limitations have also made it difficult to amplify large fragments. Additionally, HCV quasispeices exists in each individual. It is obviously that cloned cDNA may link different variants through the short RT-PCR fragments. In this study serum specimens of HCV patients and their clinical data are collected to establish HCV repository in China. Then the amplification method of long RT-PCR of HCV was developed and the full length gemomic plasmid of HCV was constructed by utilizing the main prevalent subtype of Chinese strain.
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