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

  • Alexander KlimovEmail author
  • Amanda Balish
  • Vic Veguilla
  • Hong Sun
  • Jarad Schiffer
  • Xiuhua Lu
  • Jacqueline M. Katz
  • Kathy Hancock
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

Virus titration Hemagglutination assay Plaque assay 50% Egg infectious dose Hemagglutination inhibition assay 50% Tissue culture infectious dose Microneutralization Virus neutralization Neutralizing antibodies Influenza 

Notes

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.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Alexander Klimov
    • 1
    Email author
  • Amanda Balish
    • 1
  • Vic Veguilla
    • 1
  • Hong Sun
    • 1
  • Jarad Schiffer
    • 2
  • Xiuhua Lu
    • 1
  • Jacqueline M. Katz
    • 1
  • Kathy Hancock
    • 1
  1. 1.Influenza Division, National Center for Immunization and Respiratory DiseasesCenters for Disease Control and PreventionAtlantaUSA
  2. 2.Division of Bacterial Diseases, National Center for Immunization and Respiratory DiseasesCenters for Disease Control and PreventionAtlantaUSA

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