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Evaluation of a new hepatitis C virus sequencing assay as a routine method for genotyping
Author(s) -
Ansaldi Filippo,
Torre Francesco,
Bruzzone Bianca Marisa,
Picciotto Antonino,
Crovari Pietro,
Icardi Giancarlo
Publication year - 2001
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/1096-9071(200101)63:1<17::aid-jmv1002>3.0.co;2-2
Subject(s) - genotype , genotyping , virology , subtyping , hepatitis c virus , biology , amplicon , typing , hepacivirus , flaviviridae , polymerase chain reaction , virus , medicine , genetics , gene , computer science , programming language
The determination of HCV genotypes, subtypes and isolates has been helpful in understanding the evolution and the epidemiology of the virus, and is an important factor in the pre‐treatment evaluation. A new simpler and automated sequencing based system has been developed recently, the Visible Genetics TruGene™ Hepatitis C Assay. The aim of the study was to compare this new genotyping assay with reverse hybridization based Innogenetics INNO‐LiPA HCV II assay that is used most commonly. Eighty‐eight HCV‐RNA positive patients were enrolled and divided in four groups: 26 hemodialysed patients, 30 untreated patients with chronic HCV hepatitis, 12 IFN non‐responder patients with chronic HCV hepatitis, 20 asymptomatic HCV positive subjects. The 5′‐UTR region was amplified by RT‐PCR and the nucleotide sequences determined by the TruGene™ assay. In parallel, the amplicons were also tested by INNO‐LiPA. Concordant results were obtained in 80 out of 88 cases (90.9%). The new assay allowed to genotype 2 samples not typed by LiPA as 1b and 2a/c. The new system also allowed the subtyping of 3 untypable samples, classified as genotype 1 by INNO‐LiPA, as genotype 1b (1 sample) and, as genotype 4 (2 samples). The difference between these genotype 4 isolates and the closest genotype 1 isolate was 6 nucleotides. One LiPA genotype 1a sample was typed as 1b and 2 genotype 1b samples were all typed as 1a by the sequence analysis. In conclusion, the new assay is a sensitive and rapid method that is suitable for accurate large‐scale genotyping. J. Med. Virol. 63:17–21, 2001. © 2001 Wiley‐Liss, Inc.