Rapid genotyping of hepatitis C virus RNA-isolates obtained from patients residing in western Europe.
Two rapid genotyping methods for hepatitis C virus (HCV), the line probe assay (Inno-LiPA) and the subtype-specific core amplification system [Okamoto et al., (1992b) Journal of General Virology 73:673-679], were applied to 58 HCV isolates which were typed as type 1 (n = 37) and type 2 (n = 21) by sequence analysis of the 5' untranslated region (5'UTR). The line probe assay targets the 5'UTR and recognized 12 subtype 1a, 25 subtype 1b, 18 subtype 2a, 2 subtype 2b and 1 subtype 2d in accordance with sequence analysis of this region. Subtype-specific core amplification revealed 7 discrepancies among the 37 type 1 isolates when compared to LiPA. A different subtype was observed in 3 isolates (1a versus 1b), 2 isolates remained untyped and 2 isolates showed a coinfection of subtype 1a and 1b. The first 5 discrepancies were confirmed by sequence analysis of the core region whereas the coinfection could not be confirmed. Of the 21 type 2 isolates only one could be typed by subtype-specific core amplification. HCV RNA was detected in all 21 cases after the general first round of polymerase chain reaction (PCR). Direct sequencing of the core region indicated sequence variation as a source of failure. It is concluded that LiPA results are conclusive for typing of HCV. However, LiPA is hampered occasionally for subtyping by lack of subtype-specific sequence variation in 5'UTR. Subtyping results by subtype-specific core amplification were accurate. However, it seems that this assay is not suitable for the identification of genotype 2 isolates that circulate in patients living in Western Europe.
Kleter
G E
GE
Department of Virology, Erasmus University Rotterdam, The Netherlands.
van Doorn
L J
LJ
Stuyver
L
L
Maertens
G
G
Brouwer
J T
JT
Schalm
S W
SW
Heijtink
R A
RA
Quint
W G
WG
D000328:Adult; D000368:Aged; D001483:Base Sequence; D001530:Belgium; D004279:DNA, Viral; D005060:Europe; D005838:Genotype; D016174:Hepacivirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008875:Middle Aged; D008969:Molecular Sequence Data; D009426:Netherlands; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000097:blood*; D012689:Sequence Homology, Nucleic Acid
D004279:DNA, Viral; D012367:RNA, Viral