title: A replicon-based shuttle vector system for assessing the phenotype of HCV NS5B polymerase genes isolated from patient populations.
abstract: Hepatitis C virus (HCV) replicon-based shuttle vectors that permit phenotypes of NS5B polymerase genes from a large number of patient isolates to be rapidly assessed when transiently expressed in cultured cells were designed. When used to test responses to an inhibitor of HCV RNA-dependent RNA polymerase, IC(50) values for inhibition covered a several hundred-fold range among 47 patient samples tested. This observation highlights the variability that can be found by testing isolates derived from HCV-infected subjects. Partial suppression with a polymerase inhibitor of the most sensitive species permitted detection of minor quasispecies that were 7-200-fold more resistant than the bulk population in approximately half of the samples. Sequence analysis showed a wide range of amino acid changes not detected by conventional selection methods using laboratory-derived strains. This approach provides a means to assess variation in antiviral efficacy, and to predict possible responses in a clinical setting.
authors.affiliation: Abbott Laboratories, Global Pharmaceutical Research and Development, Department R4CQ, AP52N, 200 Abbott Park Road, Abbott Park, IL 60064, USA. timothy.middleton@abbott.com
authors.forename: Tim
authors.initials: T
authors.lastname: Middleton
authors.forename: Yupeng
authors.initials: Y
authors.lastname: He
authors.forename: Tami
authors.initials: T
authors.lastname: Pilot-Matias
authors.forename: Rakesh
authors.initials: R
authors.lastname: Tripathi
authors.forename: Ben Hock
authors.initials: BH
authors.lastname: Lim
authors.forename: Andrew
authors.initials: A
authors.lastname: Roth
authors.forename: Chih-Ming
authors.initials: CM
authors.lastname: Chen
authors.forename: Gennadiy
authors.initials: G
authors.lastname: Koev
authors.forename: Teresa I
authors.initials: TI
authors.lastname: Ng
authors.forename: Preethi
authors.initials: P
authors.lastname: Krishnan
authors.forename: Ron
authors.initials: R
authors.lastname: Pithawalla
authors.forename: Rubina
authors.initials: R
authors.lastname: Mondal
authors.forename: Tatyana
authors.initials: T
authors.lastname: Dekhtyar
authors.forename: Liangjun
authors.initials: L
authors.lastname: Lu
authors.forename: Hongmei
authors.initials: H
authors.lastname: Mo
authors.forename: Warren M
authors.initials: WM
authors.lastname: Kati
authors.forename: Akhteruzzaman
authors.initials: A
authors.lastname: Molla
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology; D024882:Drug Resistance, Viral; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D005838:Genotype; D016174:Hepacivirus / Q000187:drug effects / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D006526:Hepatitis C / Q000821:virology*; D006801:Humans; D010641:Phenotype; D010957:Plasmids; D012367:RNA, Viral / Q000302:isolation & purification; D012324:RNA-Dependent RNA Polymerase / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000302:isolation & purification; D012093:Replicon; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D000998:Antiviral Agents; D004791:Enzyme Inhibitors; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase