As. Muerhoff et al., IDENTIFICATION OF CONSERVED NUCLEOTIDE-SEQUENCES WITHIN THE GB VIRUS-C 5'-UNTRANSLATED REGION - DESIGN OF PCR PRIMERS FOR DETECTION OF VIRAL-RNA, Journal of virological methods, 62(1), 1996, pp. 55-62
Citations number
12
Categorie Soggetti
Virology,"Biochemical Research Methods","Biothechnology & Applied Migrobiology
Recently, the discovery of a new human RNA virus, GB virus C (GBV-C),
was reported. GBV-C was isolated from the serum of a West African indi
vidual using degenerate oligonucleotide PCR primers designed from a co
nsensus sequence of the NS3 helicase genes of hepatitis C virus (HCV),
GBV-A, and GBV-B. Seven other individuals were shown to be infected w
ith GBV-C via RT-PCR using these primers. Subsequently, degenerate PCR
primers based upon a consensus sequence of the eight original isolate
s were designed. These primers were shown to be superior to the origin
al set. However, since they were derived from a region of the viral ge
nome exhibiting up to 17% nucleotide sequence divergence, mismatch bet
ween the primers and template may result in an underestimation of the
true GBV-C prevalence. To overcome this potential problem, we obtained
the sequences at the 5'-untranslated region (UTR) of the GBV-C genome
from 35 infected individuals and identified regions of high sequence
conservation among the isolates. We describe the design and testing of
PCR primers derived from conserved sequences within the 5'-UTR of the
GBV-C genome. These primers were shown to be as effective as the heli
case-derived primers in detecting GBV-C RNA in human sera.