Detection of hepatitis C virus RNA in liver tissues by an in situ hybridization technique.
Hepatitis C virus (HCV) infection of cells in liver tissues was determined by detecting HCV RNA by an in situ hybridization technique using synthetic oligonucleotide probes derived from the 5'-non-coding and core regions of HCV genome. Aggregated silver grains indicating hybridization with HCV RNA were observed over the nuclei as well as the cytoplasm of hepatocytes with none on non-parenchymal cells. The specificity of the hybridization was confirmed by absence of autoradiographic signals after ribonuclease predigestion, addition of an excess of non-labeled probes, or application of an M 13 probe. The hepatocytes with HCV RNA-positive signals were scattered in the periportal and mediolobular zones of liver lobules rather than in the pericentral zones. Fifteen out of 33 biopsy specimens from patients with chronic HCV infection studied had the HCV RNA-positive hepatocytes. These cells were more frequently detected in specimens with advanced periportal, bridging and intralobular necrosis but showed no correlation with the extent of inflammatory cell infiltration. These findings suggest a close correlation between the detection of HCV RNA in hepatocytes and advanced necrosis of the specimens.
First Department of Medicine, Osaka University Medical School, Japan.
Y
Y
Haruna
N
N
Hayashi
N
N
Hiramatsu
T
T
Takehara
H
H
Hagiwara
Y
Y
Sasaki
A
A
Kasahara
H
H
Fusamoto
T
T
Kamada
D000328:Adult; D000368:Aged; D001483:Base Sequence; D001706:Biopsy; D005260:Female; D016174:Hepacivirus / Q000302:isolation & purification*; D006526:Hepatitis C / Q000382:microbiology* / Q000473:pathology; D006801:Humans; D017403:In Situ Hybridization; D008099:Liver / Q000382:microbiology* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012367:RNA, Viral / Q000032:analysis*
D015345:Oligonucleotide Probes; D012367:RNA, Viral