Amplification of GB virus-C/hepatitis G virus RNA with primers from different regions of the viral genome.
GB virus-C/hepatitis G virus (GBV-C/HGV) is a newly identified RNA virus. The aim of the study was to compare three primer pairs from the 5' untranslated region (5'UTR), envelope region 2 (E 2) and nonstructural region 3 (NS 3) of GBV-C/HGV genome for their ability to detect GBV-C/HGV RNA by polymerase chain reaction (PCR) assays. By using PCR with primers from different regions of the viral genome, serum GBV-C/HGV RNA was assayed in 200 at-risk individuals. The sensitivity of this assay was assessed by a titration experiment, and nucleotide sequences of the amplified products were determined directly. Of 200 serum samples, 43 (21.5%) were positive for GBV-C/HGV RNA with at least one of the primer pairs. The positive rates by 5'UTR, NS 3, and E 2 primers were 100%, 98%, and 84%, respectively, and the sensitivity of PCR assays using 5'UTR primers was 10 to 100 times more likely to detect GBV-C/HGV RNA than that of NS 3 and E 2 primers. The average homology of amplified targets to the prototype HGV genome was 89%, 80%, and 85% and the similarity between each amplified target was up to 100%, 90%, and 92% in the 5'UTR, E 2, and NS 3 regions, respectively. Therefore, the 5'UTR of GBV-C/HGV genome is highly conserved and primers deduced from this region can provideva sensitive and specific PCR assay for GBV-C/HGV RNA.
Kao
J H
JH
Department of Internal Medicine, National Taiwan University Hospital, National Taiwan University College of Medicine, Taipei.
Chen
P J
PJ
Chen
W
W
Hsiang
S C
SC
Lai
M Y
MY
Chen
D S
DS
D001483:Base Sequence; D017931:DNA Primers; D004279:DNA, Viral; D018067:Flaviviridae / Q000235:genetics*; D016679:Genome, Viral; D006525:Hepatitis, Viral, Human / Q000821:virology*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012367:RNA, Viral; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004279:DNA, Viral; D012367:RNA, Viral