Avian influenza virus isolation, propagation and titration in embryonated chicken eggs

Authors: Spackman, Erica; KILLIAN, MARYLEA - ANIMAL AND PLANT HEALTH INSPECTION SERVICE (APHIS)

Submitted to: Animal Influenza Virus
Publication Type: Book / Chapter
Publication Acceptance Date: 5/1/2019
Publication Date: 3/27/2020
Citation: Spackman, E., Killian, M. 2020. Avian influenza virus isolation, propagation and titration in embryonated chicken eggs. In: Spackman, E., editor. Animal Influenza Virus Method and Protocols. 3rd Edition. Methods in Molecular Biology (volume 2123). New York, NY: Humana. p. 149-164. https://doi.org/10.1007/978-1-0716-0346-8_11.
DOI: https://doi.org/10.1007/978-1-0716-0346-8

Interpretive Summary:

Technical Abstract: Avian influenza virus and some mammalian influenza A viruses can be isolated, propagated, and titrated in embryonated chicken eggs (ECE). Most any sample type 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 (see Chapter X). Culture in ECE 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, antigen immunoassay, agar gel immunodiffusion assay, or RT-PCR. Finally, ECE may be used to propagate and titrate avian influenza virus.