In situ detection of hepatitis C virus RNA in liver tissue using a digoxigenin-labeled probe created during a polymerase chain reaction.
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.
Cho
S W
SW
Department of Internal Medicine, Soon Chun Hyang University Hospital, Seoul, Korea.
Hwang
S G
SG
Han
D C
DC
Jin
S Y
SY
Lee
M S
MS
Shim
C S
CS
Lee
D W
DW
Lee
H B
HB
D000328:Adult; D000368:Aged; D001483:Base Sequence; D004396:Coloring Agents / Q000494:pharmacology; D017931:DNA Primers; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis; D004076:Digoxigenin / Q000494:pharmacology; D005260:Female; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006526:Hepatitis C / Q000473:pathology / Q000821:virology*; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D008099:Liver / Q000473:pathology / Q000821:virology*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis*; D012680:Sensitivity and Specificity
D004396:Coloring Agents; D017931:DNA Primers; D015342:DNA Probes; D004279:DNA, Viral; D012367:RNA, Viral; D004076:Digoxigenin