In Situ Localization of PCR Amplified cDNA for the Diagnosis of HCV Infection in Egyptian Liver Biopsies

Journal of The Egyptian National Cancer Institute

Volume 12 - Number 3

Article Type: Original Article
Abstract: Hepatitis C virus (HCV), a major cause of chronic liv-er disease, is highly prevalent in Egypt. The morpho-logical criteria for diagnosis of HCV in liver tissue are suggestive, while the definite diagnosis relies on direct de-tection of HCV RNA. We attempted at localizing HCV gnomic RNA in for-malin fixed paraffin embedded liver tissue sections using the reverse transcription polymerase chain reaction in situ hybridization technique (RT in situ - PCR) versus standard in situ hybridization (ISH). Liver biopsies from 43 patients with clinical and his-tological evidence suggestive of HCV infection were in-cluded. PCR amplified viral complimentary DNA (cDNA) was detected in liver hepatocytes by in situ hybridization with a biotin labeled probe, in 32 cases, 32/43 (74.4%). The signal was seen as strong perinuclear reaction with occasional cytoplasmic reaction. From the 32 tissues found positive for HCV by RT in situ PCR, only 9 were positive using standard in situ hybridization, 9/32 (28.1%), with a weaker signal obtained. The present study recommends applying RT in situ PCR as a sensitive method for the confident diagnosis of HCV in liver biopsies that might help to assess morpho-logic criteria. The objective diagnosis of HCV by RT in situ PCR could limit therapeutic lines to positive cases.