Title: A cellular replicon-based phenotyping assay to determine susceptibility of hepatitis C virus clinical isolates to NS3/4A protease inhibitors
Authors: Vijgen, Leen
Verbeeck, Jannick
Van Kerckhove, Barbara
Berke, Jan Martin
Koletzki, Diana
Fanning, Gregory
Lenz, Oliver #
Series Editors: Walker, John M
Issue Date: 2013
Publisher: Humana Press
Series Title: Methods in Molecular Biology vol:1030
Host Document: Antiviral Methods and Protocols edition:2nd pages:105-117
Article number: 9
Abstract: A hepatitis C virus (HCV) replicon-based protease phenotyping assay has been developed that allows determining the susceptibility of a patient’s HCV protease sequence to HCV protease inhibitors. In brief, HCV protease sequences amplified from clinical samples are cloned in a transient HCV genotype 1b replicon backbone, containing a luciferase reporter gene. These protease chimeric replicons are replication-competent when electroporated into susceptible Huh7-Lunet cells. Replication can be quantified by measuring the enzymatic activity of the luciferase protein. This assay is reproducible and robust, and has a high overall success rate for determining the phenotypic susceptibility of HCV genotype 1a and 1b patient-derived protease domains to HCV protease inhibitors. In addition, the HCV genotype 1b protease shuttle backbone also supports efficient replication of HCV genotype 4 protease sequences.
ISBN: 978-1-62703-484-8
ISSN: 1064-3745
Publication status: published
KU Leuven publication type: IHb
Appears in Collections:Non-KU Leuven Association publications
# (joint) last author

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