Avian Influenza Virus Isolation, Propagation, and Titration in Embryonated Chicken Eggs

  • Erica SpackmanEmail author
  • Mary Lea Killian
Part of the Methods in Molecular Biology book series (MIMB, volume 2123)


Avian influenza virus and some mammalian influenza A viruses can be isolated, propagated, and titrated in embryonated chicken eggs (ECEs). Most sample types can be accommodated in ECE culture with appropriate processing. Isolation may also be accomplished in cell culture, and if a mammalian lineage influenza A is expected to be in an avian sample, for example swine influenza in turkey specimens, mammalian cell may be preferable. Culture in ECEs is highly sensitive but is not specific for influenza A, which may be an advantage because a sample may be screened for several agents at once. Once an agent is isolated in culture, the presence of influenza viruses can be confirmed by hemagglutination inhibition assay, antigen immunoassay, agar gel immunodiffusion assay, or RT-PCR. Finally, ECEs may be used to propagate and titrate an avian influenza virus.

Key words

Avian influenza Virus isolation Virus culture Embryonated chicken eggs 


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Copyright information

© This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply. 2020

Authors and Affiliations

  1. 1.Exotic and Emerging Avian Viral Diseases Unit, US National Poultry Research CenterUS Department of Agriculture, Agricultural Research ServiceAthensUSA
  2. 2.Avian Viruses Section, Diagnostic Virology Laboratory, National Veterinary Services Laboratories, US Department of AgricultureAmesUSA

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