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.
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References
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Vijgen, L. et al. (2013). A Cellular Replicon-Based Phenotyping Assay to Determine Susceptibility of Hepatitis C Virus Clinical Isolates to NS3/4A Protease Inhibitors. In: Gong, E. (eds) Antiviral Methods and Protocols. Methods in Molecular Biology, vol 1030. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-484-5_9
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DOI: https://doi.org/10.1007/978-1-62703-484-5_9
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-483-8
Online ISBN: 978-1-62703-484-5
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