Skip to main content
SpringerLink
Log in

Influenza Virus Titration, Antigenic Characterization, and Serological Methods for Antibody Detection

  • Protocol
  • First Online:
Influenza Virus

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

Abstract

This chapter describes some commonly used methods of influenza virus titration, antigenic characterization, and serological methods by antibody detection. These methods are essential not only for virus characterization but also for identifying new antigenic variants, vaccine strain selection, and sero-epidemiologic studies of influenza virus transmission and prevalence. Virus titration methods such as the hemagglutination assay, 50% egg or tissue culture infectious dose, and plaque assay are employed to determine the amount of virus particles in a sample. The hemagglutination inhibition assay is a reliable, relatively simple and inexpensive technique to antigenically characterize isolates of influenza viruses. Serological methods such as virus neutralization and hemagglutination inhibition are the fundamental tools used in sero-epidemiologic studies of influenza virus transmission and prevalence and in the evaluation of vaccine immunogenicity. While serological methods rarely yield an early diagnosis of acute influenza virus infection, well-timed, paired acute, and convalescent serum samples may establish the diagnosis of a recent influenza infection even when attempts to detect the virus are negative.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
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. Hirst, G.K. (1941) Agglutination of red cells by amniotic fluid of chick embryos infected with influenza virus. Science 94, 22–23.

    Article  PubMed  CAS  Google Scholar 

  2. Miller, G.L. and W.M. Stanley. (1944) Quantitative aspects of the red blood cell agglutination test for influenza virus. J. Exp. Med. 79, 185–95.

    Article  PubMed  CAS  Google Scholar 

  3. Miller, G.L. (1965) Improved measurement of influenza virus hemagglutinin titer. J. of Immuno. 95, 336–344.

    CAS  Google Scholar 

  4. Reed, L.J. and H. Muench. (1938) A simple method of estimating fifty per cent endpoints. Am. J. Hyg. 27, 493–497.

    Google Scholar 

  5. Tobita, K., A. Sugiura, C. Enomoto, and M. Furuyama. (1975) Plaque assay and primary isolation of inflyenza A viruses in an established line of canine kidney cells (MDCK) in the presence of trypsin. Med. Micro. Immuno. 162, 9–14.

    Article  CAS  Google Scholar 

  6. Tobita, K. (1975) Permanent canine kidney (MDCK) cells for isolation and plaque assay of influenza B viruses. Med. Micro. Immuno. 162, 23–7.

    Article  CAS  Google Scholar 

  7. Hirst, G.K. (1942) The quantitative determination of influenza virus and antibodies by means of red cell agglutination. J. Exp. Med. 75, 47–64.

    Google Scholar 

  8. Salk, J. E. (1944) Simplified procedure for titrating hemagglutinating capacity of influenza virus and the corresponding antibody. J. Immun. 49, 87.

    Google Scholar 

  9. Rowe, T., Abernathy, R. A., Hu-Primmer, J., Thompson, W.W., Lu, X., Lim, W., Fukuda, K., Cox, N. J., and Katz, J. M. (1999) Detection of antibody to avian influenza A (H5N1) virus in human serum by using a combination of serologic assays. J. Clin. Micro. 37, 937–343.

    CAS  Google Scholar 

  10. Stephenson, I., Das, R. G., Wood, J. M., and Katz, J.M. (2007) Comparison of neutralizing antibody assays for detection of antibody to influenza A/H3N2 viruses: An international collaborative study. Vaccine 25, 4056–4063.

    Article  PubMed  CAS  Google Scholar 

  11. Stephenson, I., Heath, A., Major, D., Newman, R. W., Hoschler, K., Junzi, W., Katz, J. M., Weir, J. P., Zambon, M. C., and Wood, J. M. (2009) Reproducibility of serologic assays for influenza virus A (H5N1). EID 15, 1250–1259.

    Google Scholar 

  12. Gaush, C.R., Hard, W. L., and Smith, T. F. (1966) Characterization of an established line of canine kidney cells (MDCK). Proc. Soc. Exp. Biol. Med. 122, 931–935.

    PubMed  CAS  Google Scholar 

  13. Medeiros, R. N. Escriou, N. Naffakh, JC Manuguerra and S. Van der Werf. (2001). Hemagglutination residues of recent human A (H3N2) influenza viruses that contribute to the inability to agglutinate chicken erythrocytes. Virol. 289, 74–85.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The microneutralization protocol presented here is based on the work of Thomas Rowe and others who developed the original microneutralization protocol for avian influenza H5N1 viruses. Thanks is expressed to Yaohui Bai and Li Cronin for their work determining the number of MDCK cells needed to obtain flasks of cells at the appropriate confluency for the microneutralization assay.

Author information

Authors and Affiliations

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

    Alexander Klimov, Amanda Balish, Vic Veguilla, Hong Sun, Xiuhua Lu, Jacqueline M. Katz & Kathy Hancock

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

    Jarad Schiffer

Authors
  1. Alexander Klimov
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Amanda Balish
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Vic Veguilla
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hong Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jarad Schiffer
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xiuhua Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jacqueline M. Katz
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Kathy Hancock
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Alexander Klimov .

Editor information

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

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Klimov, A. et al. (2012). Influenza Virus Titration, Antigenic Characterization, and Serological Methods for Antibody Detection. 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_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-621-0_3

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-620-3

  • Online ISBN: 978-1-61779-621-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation