A novel method for the measurement of hepatitis C virus infectious titres using the IncuCyte ZOOM and its application to antiviral screening

Journal of Virological Methods
Hazel StewartMark Harris

Abstract

Hepatitis C virus (HCV) is a significant human pathogen infecting 3% of the world population. An infectious molecular clone capable of replicating and releasing infectious virions in cell culture has only been available since 2005, leaving a significant knowledge gap concerning post-RNA replication events such as particle assembly, trafficking and release. Thus, a fast, efficient and accurate method of measuring infectious viral titres is highly desirable. Current methods rely upon manual counting of infected cell foci and so are both labour-intensive and susceptible to human error. Here, we report a novel protocol, which utilises the IncuCyte ZOOM instrument and related software to accurately count infected cells and extrapolation of this data to produce an infectious titre, reported as infectious units per millilitre (IU/mL). This method reduces cost, time and error in experiments. We also demonstrate that this approach is amenable to high-throughput compound screening, thereby expediting the identification of novel antivirals.

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Citations

Jun 22, 2016·The Journal of General Virology·Eleni-Anna LoundrasNicola J Stonehouse
Dec 3, 2017·The Journal of Immunology : Official Journal of the American Association of Immunologists·Sarthak GuptaMariana J Kaplan
Nov 16, 2017·The Journal of General Virology·Niluka GoonawardaneMark Harris
Jan 21, 2018·PLoS Pathogens·Chunhong YinMark Harris
Mar 23, 2018·International Journal of Molecular Sciences·Margarita-Maria PanouAndrew Macdonald
Apr 2, 2020·Antiviral Research·Margarita-Maria PanouAndrew Macdonald
Mar 12, 2021·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Rongxin FuGuoliang Huang

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Methods Mentioned

BETA
in vitro transcription
transfection

Software Mentioned

GraphPad
Prism
IncuCyte ZOOM
IncuCyte

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