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    Fluorescence-based Neuraminidase Inhibition Assay to Assess the Susceptibility of Influenza Viruses to The Neuraminidase Inhibitor Class of Antivirals

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    Author
    Leang, S-K; Hurt, AC
    Date
    2017-04-01
    Source Title
    Journal of Visualized Experiments
    Publisher
    JOURNAL OF VISUALIZED EXPERIMENTS
    University of Melbourne Author/s
    Hurt, Aeron
    Affiliation
    Melbourne School of Population and Global Health
    Metadata
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    Document Type
    Journal Article
    Citations
    Leang, S. -K. & Hurt, A. C. (2017). Fluorescence-based Neuraminidase Inhibition Assay to Assess the Susceptibility of Influenza Viruses to The Neuraminidase Inhibitor Class of Antivirals. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2017 (122), https://doi.org/10.3791/55570.
    Access Status
    Open Access
    URI
    http://hdl.handle.net/11343/259106
    DOI
    10.3791/55570
    Abstract
    The neuraminidase (NA) inhibitors are the only class of antivirals approved for the treatment and prophylaxis of influenza that are effective against currently circulating strains. In addition to their use in treating seasonal influenza, the NA inhibitors have been stockpiled by a number of countries for use in the event of a pandemic. It is therefore important to monitor the susceptibility of circulating influenza viruses to this class of antivirals. There are different types of assays that can be used to assess the susceptibility of influenza viruses to the NA inhibitors, but the enzyme inhibition assays using either a fluorescent substrate or a chemiluminescent substrate are the most widely used and recommended. This protocol describes the use of a fluorescence-based assay to assess influenza virus susceptibility to NA inhibitors. The assay is based on the NA enzyme cleaving the 2'-(4-Methylumbelliferyl)-α-D-N-acetylneuraminic acid (MUNANA) substrate to release the fluorescent product 4-methylumbelliferone (4-MU). Therefore, the inhibitory effect of an NA inhibitor on the influenza virus NA is determined based on the concentration of the NA inhibitor that is required to reduce 50% of the NA activity, given as an IC50 value.

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