title: High-throughput titration of luciferase-expressing recombinant viruses.
abstract: Standard plaque assays to determine infectious viral titers can be time consuming, are not amenable to a high volume of samples, and cannot be done with viruses that do not form plaques. As an alternative to plaque assays, we have developed a high-throughput titration method that allows for the simultaneous titration of a high volume of samples in a single day. This approach involves infection of the samples with a Firefly luciferase tagged virus, transfer of the infected samples onto an appropriate permissive cell line, subsequent addition of luciferin, reading of plates in order to obtain luminescence readings, and finally the conversion from luminescence to viral titers. The assessment of cytotoxicity using a metabolic viability dye can be easily incorporated in the workflow in parallel and provide valuable information in the context of a drug screen. This technique provides a reliable, high-throughput method to determine viral titers as an alternative to a standard plaque assay.
authors.affiliation: Center for Innovative Cancer Research, Ottawa Hospital Research Institute; Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Garcia
authors.affiliation: Center for Innovative Cancer Research, Ottawa Hospital Research Institute; Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa.
authors.forename: Ramya
authors.initials: R
authors.lastname: Krishnan
authors.affiliation: Center for Innovative Cancer Research, Ottawa Hospital Research Institute; Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa.
authors.forename: Colin
authors.initials: C
authors.lastname: Davis
authors.affiliation: Center for Innovative Cancer Research, Ottawa Hospital Research Institute; Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa.
authors.forename: Cory
authors.initials: C
authors.lastname: Batenchuk
authors.affiliation: Center for Innovative Cancer Research, Ottawa Hospital Research Institute; Faculty of Medicine, University of Ottawa.
authors.forename: Fabrice
authors.initials: F
authors.lastname: Le Boeuf
authors.affiliation: Center for Innovative Cancer Research, Ottawa Hospital Research Institute; Faculty of Medicine, University of Ottawa.
authors.forename: Hesham
authors.initials: H
authors.lastname: Abdelbary
authors.affiliation: Center for Innovative Cancer Research, Ottawa Hospital Research Institute; Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa; jsdiallo@ohri.ca.
authors.forename: Jean-Simon
authors.initials: JS
authors.lastname: Diallo
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D057166:High-Throughput Screening Assays / Q000379:methods*; D049409:Luciferases, Firefly / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D019076:Transgenes; D014709:Vero Cells; D018116:Vesiculovirus / Q000201:enzymology* / Q000235:genetics; D014776:Virus Cultivation / Q000379:methods*
chemical_list: D049409:Luciferases, Firefly