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INTRODUCTION
HCV (hepatitis C virus) infects nearly 3% of the population worldwide and has emerged as a major causative agent of liver disease, resulting in acute and chronic infections that can lead to fibrosis, cirrhosis and hepato-cellular carcinoma. Current laboratory diagnosis of HCV is based on specific antibody detection (anti-hepatitis C virus-anti-HCV) in serum. Determination of HCV RNA concentrations reduces the pre-seroconversion period in the diagnosis of HCV infection and supports management of interferon alpha-based therapies. As HCV replicates in the liver cells of these patients, detection and localization of HCV RNA in liver tissue are vital for diagnostic purposes and clinical management of such patients, as well as for the elucidation of viro-pathological mechanisms.
MATERIALS AND METHODS
A total of 10 biopsy samples diagnosed for liver cirrhosis that were negative for the presence of anti-HCV and serum HCV RNA were undertaken for analyzing the presence of viral nucleic acid in liver tissues. The diagnosis of liver cirrhosis was made on the grounds of histology and/or laparoscopy. Since no defined cause of liver cirrhosis was identified the etiology of liver cirrhosis was classified as cryptogenic. Qualitative screening for HCV was done through ELISA while the nucleic acid analysis was performed through Light Cycler 2.0 (minimum detection limit 10 copies/ml). The detection of HCV RNA in liver tissue biopsies was performed by following standard protocol of HCV detection kit (Shenzhen PG Biotech) with modifications using Light Cycler 2.0.
RESULTS
It was observed that all the 10 samples were negative for the anti-HCV and serum HCV RNA. While the quantitative detection in liver biopsies following the modified method showed the presence of HCV RNA in 3 samples out of the 10 studied.
CONCLUSION
The observed results indicate that the Light Cycler 2.0 following the modified technique described here may constitute a reliable method of quantitative detection and localization of HCV in tissue, since it allows identification of specific regions of the viral genome with high confidence. It may be an important investigative tool to study "serosilent" HCV infection in cryptogenic cases of liver. Further studies, comprising a larger sample are under development in order to assess possible association between the presence of HCV-RNA in liver and clinical manifestations, histopathological status, viral genotypes, and even in patients without evidence of circulating HCV.