In situ detection of hepatitis C virus RNA in liver tissue using a digoxigenin-labeled probe created during a polymerase chain reaction.

Abstract

The cellular localization of hepatitis C virus (HCV) RNA in liver tissue was studied by nonisotopic in situ hybridization using a digoxigenin-labeled cDNA probe created during a polymerase chain reaction on samples from 16 patients with chronic HCV infection. Hybridization signals were recognized in the cytoplasm of the hepatocytes, and a few hepatocytes had hybridization signals in the nucleus as well. HCV RNA positive hepatocytes were found in 1 of 9 patients with chronic persistent hepatitis, 2 of 5 patients with chronic active hepatitis, and in each of 2 patients with chronic active hepatitis and cirrhosis. Positive signals were found in many hepatocytes within the lobule in liver sections of patients with advanced chronic active hepatitis. A number of HCV RNA positive hepatocytes were found in nodules, but not in the area of fibrosis. On the other hand, positive signals were found in a few hepatocytes scattered in the lobule in a patient with chronic persistent hepatitis. The mean ALT levels in the patients with positive signal (175.6 +/- 44.2 U/L) were significantly higher than in those without a signal (70.27 +/- 16.1 U/L) (P < 0.05). The findings suggest that a larger amount of HCV may be present during the advanced than during the early stages of type C hepatitis and nonisotopic in situ hybridization using a digoxigenin-labeled HCV cDNA probe created during a polymerase chain reaction deserves wider application for the detection of HCV replication in specimens.