Establishment of a highly specific detection system for GB virus C (GBV-C) minus-strand RNA.
Although the clinical relevance of GB virus C (GBV-C) is still elusive, this virus has been found with high prevalence in several groups of patients with liver disease. As was shown for hepatitis C virus (HCV), minus-strand RNA is supposed to function as a replicative intermediate. We have established a reliable and sensitive detection system for GBV-C minus-strand RNA based on nested RT-PCR (reverse transcription-polymerase chain reaction) with a tagged primer system. Sensitivity and specificity was extensively tested using in-vitro transcribed GBV-C sequences and genomic viral RNA. Specificity of the amplified fragments was proven by Southern blot hybridization. Using this detection system, we found the presence of GBV-C minus-strand RNA in 6/41 (14.6%) sera of GBV-C infected or GBV-C/HCV coinfected patients. No correlation with virological parameters such as amount of GBV-C plus-strand RNA, genotype or titer of HCV could be detected.
Seipp
S
S
Institute of Zoology, University of Heidelberg, Germany. stefanie.seipp@www.zoo.uni-heidelberg.de
Goeser
T
T
Theilmann
L
L
Kallinowski
B
B
D000328:Adult; D015139:Blotting, Southern; D005260:Female; D018067:Flaviviridae / Q000235:genetics / Q000302:isolation & purification*; D005814:Genes, Viral / Q000235:genetics; D005819:Genetic Markers; D006526:Hepatitis C / Q000097:blood / Q000821:virology; D006525:Hepatitis, Viral, Human / Q000097:blood / Q000821:virology; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
D005819:Genetic Markers; D012367:RNA, Viral