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The Fluorescence Neuraminidase Inhibition Assay: A Functional Method for Detection of Influenza Virus Resistance to the Neuraminidase Inhibitors

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Influenza Virus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 865))

Abstract

Neuraminidase inhibitors (NAIs) are presently the only effective antiviral drugs for treatment and chemoprophylaxis of influenza A and B infections, due to the high prevalence of resistance to the adamantane class of drugs among influenza A(H3N2) and A(H1N1) viruses, including the 2009 pandemic H1N1 strain. The limited pharmaceutical options currently available for control of influenza infections underscore the critical need for surveillance on NAI susceptibility of influenza viruses circulating globally. This chapter describes the fluorescent neuraminidase (NA) inhibition (NI) assay, a functional method used for assessing influenza virus susceptibility to NAIs. The IC50 (drug concentration needed to reduce the NA enzymatic activity by 50%) values generated in this assay are used to evaluate the NAI-susceptibility of test viruses relative to those of sensitive reference viruses of the same antigenic type and subtype. Test viruses with significantly elevated IC50s are further analyzed by pyrosequencing or conventional sequencing to identify known markers of NAI resistance or novel changes in the NA. The harmonization of NI assay conditions and interpretation of results across surveillance laboratories is necessary to improve NAI susceptibility testing and analysis.

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Acknowledgments

The Melbourne WHO Collaborating Centre for Reference and Research on Influenza is supported by the Australian Government Department of Health and Ageing. This work was part funded by the US Centers for Disease Control and Prevention.

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Authors and Affiliations

  1. Victorian Infectious Diseases Reference Laboratory, WHO Collaborating Centre for Reference and Research on Influenza, Melbourne, VIC, Australia

    Aeron C. Hurt

  2. Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA

    Margaret Okomo-Adhiambo & Larisa V. Gubareva

  3. WHO Collaborating Center for Surveillance, Epidemiology and Control of Influenza, Atlanta, GA, USA

    Margaret Okomo-Adhiambo & Larisa V. Gubareva

Authors
  1. Aeron C. Hurt
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  2. Margaret Okomo-Adhiambo
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  3. Larisa V. Gubareva
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Corresponding author

Correspondence to Larisa V. Gubareva .

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Editors and Affiliations

  1. School of Veterinary Medicine, Dept. Pathobiological Science, University of Wisconsin-Madison, Science Drive 575, Madison, 53711, Wisconsin, USA

    Yoshihiro Kawaoka

  2. Influenza Research Institute (IRI), Dept. Pathobiological Science, University of Wisconsin-Madison, Science Drive 575, Madison, 53711, Wisconsin, USA

    Gabriele Neumann

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The findings and conclusions of this report are those of the authors and do not necessarily represent the views of the Centers for Disease Control and Prevention.

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© 2012 Springer Science+Business Media, LLC

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Hurt, A.C., Okomo-Adhiambo, M., Gubareva, L.V. (2012). The Fluorescence Neuraminidase Inhibition Assay: A Functional Method for Detection of Influenza Virus Resistance to the Neuraminidase Inhibitors. In: Kawaoka, Y., Neumann, G. (eds) Influenza Virus. Methods in Molecular Biology, vol 865. Humana Press. https://doi.org/10.1007/978-1-61779-621-0_7

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  • DOI: https://doi.org/10.1007/978-1-61779-621-0_7

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-620-3

  • Online ISBN: 978-1-61779-621-0

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