Detection of hepatitis C virus RNA in liver tissue of chronic hepatitis C conditions by reverse transcription-polymerase chain reaction in situ hybridization [TR-PCR-ISH]

ABSTRACT

Hepatitis C virus [HCV] infection of cells on liver tissue was determined by detecting HCV RNA using reverse transcription polymerase chain reaction in situ hybridization [RT PCR ISH] technique. Twenty wedge liver biopsy specimens from patient with chronic hepatitic conditions were studied; comprising eighteen cases of chronic active hepatitis [CAH] with cirrhosis, five of which were positive for HBsAg] and the rest were diagnosed as non A-non B [NANB] and two cases of CAH with bilharzial fibrosis were also diagnosed as [NANB]. Score activity of the 20 liver biopsies was done and features favorable of HCV infection were determined. Thirteen out of the 20 liver biopsy specimens were positive for HCV RNA which was located within hepatocyte cytoplasm for all thirteen cases, in nuclei for six cases and in bile duct epithelium for two cases. HCV RNA appeared as fine granules diffusely distributed in cytoplasm or concentrated in perinuclear zone. The positive hepatocytes were particularly related to periportal and midlobular zone in relation to lymphocytic infiltrate. A high proportion of hepatocytes appeared positive with variable intersity but there was no strong association with disease activity. HCV RNA was detected in one severe case of CAH with bilharzial fibrosis and three cases positive for ABsAgs. In this study, some histopathological characteristics of HCV infection were revealed and it was concluded that direct recognition of HCV in liver biopsy is mandatory for the establishment of the cause and effect of hepatitis and using RT-PCR-ISH technique would be the most reliable method of diagnosis of hepatitis C virus because it is more sensitive and specific