In situ localization of PCR amplified CDNA for the diagnosis of HCV infection in Egyptian liver biopsies

الملخص EN

Hepatitis C virus (HCV), a major cause of chronic liver disease, is highly prevalent in Egypt.

The morphological criteria for diagnosis of HCV in liver tissue are suggestive, while the definite diagnosis relies on direct detection of HCV RNA.

We attempted at localizing HCV gnomic RNA in formalin 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 histological evidence suggestive of HCV infection were included.

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 morphologic criteria.

The objective diagnosis of HCV by RT in situ PCR could limit therapeutic lines to positive cases.