Standardization of the filovirus plaque assay for use in preclinical studies

Amy C. Shurtleff, Julia E. Biggins, Ashley E. Keeney, Elizabeth E. Zumbrun, Holly A. Bloomfield, Ana Kuehne, Jennifer L. Audet, Kendra J. Alfson, Anthony Griffiths, Gene G. Olinger, Sina Bavari

    Research output: Contribution to journalArticlepeer-review

    38 Scopus citations

    Abstract

    The filovirus plaque assay serves as the assay of choice to measure infectious virus in a cell culture, blood, or homogenized tissue sample. It has been in use for more than 30 years and is the generally accepted assay used to titrate virus in samples from animals treated with a potential antiviral therapeutic or vaccine. As these animal studies are required for the development of vaccines and therapeutics under the FDA Animal Rule, it is essential to have a standardized assay to compare their efficacies against the various filoviruses. Here, we present an evaluation of the conditions under which the filovirus plaque assay performs best for the Ebola virus Kikwit variant and the Angola variant of Marburg virus. The indicator cell type and source, inoculum volumes, length of incubation and general features of filovirus biology as visualized in the assay are addressed in terms of the impact on the sample viral titer calculations. These optimization studies have resulted in a plaque assay protocol which can be used for preclinical studies, and as a standardized protocol for use across institutions, to aid in data comparison. This protocol will be validated for use in GLP studies supporting advanced development of filovirus therapeutics and vaccines.

    Original languageEnglish (US)
    Pages (from-to)3511-3530
    Number of pages20
    JournalViruses
    Volume4
    Issue number12
    DOIs
    StatePublished - 2012

    Keywords

    • Ebola
    • Ebolavirus
    • Filovirus
    • Marburg virus
    • Marburgvirus
    • Plaque assay
    • Vero

    ASJC Scopus subject areas

    • Infectious Diseases
    • Virology

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