Skip to main content

Virological Methods for the Generation and Characterization of Influenza Viruses with Reduced Sensitivity to 4-Guanidino-Neu5Ac2en

  • Protocol
Antiviral Methods and Protocols

Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 24))

  • 566 Accesses

Abstract

The compound 4-guanidino-Neu5Ac2en (zanamivir) has been described as a selective inhibitor of the influenza virus neuraminidase (NA) (1). Viruses that are less sensitive to this inhibitor can be isolated after several passages in MDCK cells in the presence of the inhibitor. Variants isolated so far have had mutations predominantly in the hemagglutinin (HA) gene (2). Many of these mutations appear to lower the affinity of the HA for the cellular receptor, so that there is less requirement for significant NA activity for the newly synthesized progeny virus to elute. There are three reports of a mutation in the NA gene, all at the same conserved site, glu 119 (35). In this chapter, the authors describe methods for the isolation of the mutants, and for their characterization in cell culture based assays.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. von Itzstein, M., Wu, W. Y., Kok, G. B., Pegg, M. S., Dyason, J. C., Phan, T. V., Smythe, M. L., White, H. F., Oliver, S. W., Colman, P. M., Varghese, J. N., Ryan, D. M., Woods, J. M., Bethell, R. C., Hotham, V. J., Cameron, J. M., and Penn, C. R. (1993) Rational design of potent sialidase-based inhibitors of influenza virus replication. Nature 363, 418–423.

    Article  Google Scholar 

  2. McKimm-Breschkin, J. L., Blick, T. J., Sahasrabudhe, A. V., Tiong, T., Marshall, D., Hart, G. J., Bethell, R. C., and Penn, C. R. (1996) Generation and characterization of variants of the NWS/G70C influenza virus after in vitro passage in 4-amino-Neu5Ac2en and 4-guanidino-Neu5Ac2en. Antimicrob. Agents Chemother. 40, 40–46.

    CAS  PubMed  Google Scholar 

  3. Blick, T. J., Tiong, T., Sahasrabudhe, A., Varghese, J. N., Colman, P. M., Hart, G. J., Bethell, R. C., and McKimm-Breschkin, J. L. (1995) Generation and characterization of an influenza virus neuraminidase variant with decreased sensitivity to the neuraminidase-specific inhibitor 4-guanidino-Neu5Ac2en. Virology 214, 475–484.

    Article  CAS  PubMed  Google Scholar 

  4. Gubareva, L. V., Bethell, R. C., Hart, G. J., Murti, K. G., Penn, C. R., and Webster, R. G. (1996) Characterization of mutants of influenza A virus selected with the neuraminidase inhibitor 4-guanidino-Neu5Ac2en. J. Virol. 70, 1818–1827.

    CAS  PubMed  Google Scholar 

  5. Staschke, K. A., Colacino, J. M., Baxter, A. J., Air, G. M., Bansal, A., Hornback, W. J., Munroe, J. E., and Laver, W. G. (1995) Molecular basis for resistance of influenza viruses to 4-guanidino-Neu5Ac2en. Virology 214, 642–646.

    Article  CAS  PubMed  Google Scholar 

Download references

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2000 Humana Press Inc.

About this protocol

Cite this protocol

McKimm-Breschkin, J.L., Sahasrabudhe, A., Blick, T.J., Jowett, A.J., Bethell, R.C. (2000). Virological Methods for the Generation and Characterization of Influenza Viruses with Reduced Sensitivity to 4-Guanidino-Neu5Ac2en. In: Kinchington, D., Schinazi, R.F. (eds) Antiviral Methods and Protocols. Methods in Molecular Medicine™, vol 24. Humana Press. https://doi.org/10.1385/1-59259-245-7:375

Download citation

  • DOI: https://doi.org/10.1385/1-59259-245-7:375

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-561-4

  • Online ISBN: 978-1-59259-245-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics