Avian Influenza Virus Isolation and Propagation in Chicken Eggs

  • Peter R. Woolcock
Part of the Methods in Molecular Biology™ book series (MIMB, volume 436)


The avian influenza (AI) virus is usually isolated and propagated by inoculating either swab or tissue samples from infected birds into the chorioallantoic sac of embryonating chicken eggs. This is the accepted method, but occasionally an isolation may only be successful when inoculated either into the yolk sac or onto the chorioallantoic membrane of embryonating chicken eggs. Chorioallantoic fluid is harvested from eggs with dead or dying embryos and is tested for the presence of hemagglutinating antigen. If hemagglutination-positive, this indicates that the isolate may be the AI virus. The presence of the AI virus may be confirmed by either an agar gel immunodiffusion (AGID) assay, RT-PCR specific for AI virus, or a commercially available immunoassay kit specific for type A influenza. Instructions for AI virus primary isolation and propagation, preparing antigen for an AGID test, setting up an AGID test, and interpreting results are given.


avian influenza virus isolation agar gel immunodiffusion (AGID) 


  1. 1.
    1. Alexander, D. J. (2000) A review of avain influenza in different bird species. Veterinary Microbiology, 74, 3–13.CrossRefPubMedGoogle Scholar
  2. 2.
    2. Swayne, D. E. and Halvorson, D. A. (2003) Influenza in Diseases of Poultry, 11th ed. (Y. M. Saif et al., eds.). Ames: Iowa State Press, pp. 135–160.Google Scholar
  3. 3.
    3. Woolcock, P. R., McFarland, M. D., Lai, S., and Chin, R. P. (2001) Enhanced recovery of avian influenza virus isolates by a combination of chicken embryo inoculation methods. Avian Dis. 45, 1030–1035.CrossRefPubMedGoogle Scholar
  4. 4.
    4. Alexander, D. J. (2004) Newcastle Disease in Manual of Diagnostic Tests and Vaccines for Terrestrial Animals 5th ed. Office International des Epizooties, Paris, France. pp. 270–282.Google Scholar
  5. 5.
    5. Woolcock, P. R. and Cardona, C. J. (2005) Commercial immunoassay kits for the detection of influenza virus type A: Evaluation of their use with poultry. Avian Dis. 49, 477–481.CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Peter R. Woolcock
    • 1
  1. 1.California Animal Health and Food SafetyUniversity of CaliforniaDavis, FresnoUSA

Personalised recommendations