A replicon-based shuttle vector system for assessing the phenotype of HCV NS5B polymerase genes isolated from patient populations.
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.
Middleton
Tim
T
Abbott Laboratories, Global Pharmaceutical Research and Development, Department R4CQ, AP52N, 200 Abbott Park Road, Abbott Park, IL 60064, USA. timothy.middleton@abbott.com
He
Yupeng
Y
Pilot-Matias
Tami
T
Tripathi
Rakesh
R
Lim
Ben Hock
BH
Roth
Andrew
A
Chen
Chih-Ming
CM
Koev
Gennadiy
G
Ng
Teresa I
TI
Krishnan
Preethi
P
Pithawalla
Ron
R
Mondal
Rubina
R
Dekhtyar
Tatyana
T
Lu
Liangjun
L
Mo
Hongmei
H
Kati
Warren M
WM
Molla
Akhteruzzaman
A
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
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