Detection of negative strand RNA of hepatitis C virus in infected liver and serum.
The negative strand RNA of hepatitis C virus, supposed to be a replicative intermediate of the virus appears to indicate viral replication. In this study, we detected the negative strand RNA by using reverse transcription-polymerase chain reaction with RNase A digestion to degrade the remaining positive strand genomic sequence of the virus after complementary DNA (cDNA) synthesis. In vitro transcribed positive-stranded mutant RNA was not detected by this method. Sample sera and liver tissues of 16 patients with chronic hepatitis C virus infection (liver fibrosis, 1; chronic hepatitis, 13; liver cirrhosis, 2) were analysed for negative strand RNA of hepatitis C virus. The negative strand RNA sequence was detected in 15 (93%) of 16 liver tissues and in 11 (78%) of 14 sera. The study demonstrated that negative strand RNA of hepatitis C virus in serum and liver tissue could be specifically detected.
Nakagawa
H
H
First Department of Internal Medicine, Okayama University Medical School, Japan.
Shimomura
H
H
Hasui
T
T
Tsuji
H
H
Tsuji
T
T
D000328:Adult; D001483:Base Sequence; D005260:Female; D016174:Hepacivirus / Q000235:genetics* / Q000502:physiology; D006526:Hepatitis C / Q000097:blood* / Q000235:genetics*; D006801:Humans; D008099:Liver / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000097:blood / Q000302:isolation & purification*; D014779:Virus Replication
D015345:Oligonucleotide Probes; D012367:RNA, Viral