Isolation and Propagation of Coronaviruses inEmbryonated Eggs

  • James S. Guy
Part of the Methods in Molecular Biology book series (MIMB, volume 454)


The embryonated egg is a complex structure comprising an embryo and its supporting membranes (chorioallantoic, amniotic, yolk). The developing embryo and its membranes provide the diversity of cell types that are needed for successful replication of a wide variety of different viruses. Within the family Coronaviridae, the embryonated egg has been used as a host system primarily for two group 3 coronaviruses, infectious bronchitis virus (IBV) and turkey coronavirus (TCoV), but it also has been shown to be suitable for pheasant coronavirus. IBV replicates well in the embryonated chicken egg, regardless of the inoculation route; however, the allantoic route is favored as the virus replicates extensively in chorioallantoic membrane and high titers are found in allantoic fluid. TCoV replicates only in embryo tissues, within epithelium of the intestines and bursa of Fabricius; thus amniotic inoculation is required for isolation and propagation of this virus. Embryonated eggs also provide a potential host system for studies aimed at identifying other, novel coronavirus species.

Key Words

embryonated egg allantoic amniotic chicken turkey isolation propagation diagnosis detection 


  1. 1.
    Adams, N. R., and Hofstad, M. S. (1970) Isolation of transmissible enteritis agent of turkeys in avian embryos Avian Dis. 15, 426–433.CrossRefGoogle Scholar
  2. 2.
    Cavanagh, D., and Naqi, S. A. (2003) Infectious bronchitis. In: Saif, Y. M., Barnes, H. J., Fadly, A., Glisson J. R., McDougald, L.R., and Swayne, D. E. (eds.) Diseases of Poultry, 11th Ed. Iowa State University Press, Ames, pp. 101–120.Google Scholar
  3. 3.
    Gough, R. E., Cox, W. J., Winkler, C. E., Sharp, M. W., and Spackman, D. (1996) Isolation and identification of infectious bronchitis virus from pheasants Vet. Rec. 138, 208–209.PubMedCrossRefGoogle Scholar
  4. 4.
    Guy, J. S. (2003) Turkey coronavirus enteritis. In: Saif, Y. M., Barnes, H. J., Fadly, A., Glisson, J. R., McDougald, L. R., and Swayne, D. E., (eds.) Diseases of Poultry, 11th Ed. Iowa State University Press, Ames, pp. 300–308.Google Scholar
  5. 5.
    Hawkes, R. A. (1979) General principles underlying laboratory diagnosis of viral infections. In: Lennette, E. H, and Schmidt, N. J. (eds.) Diagnostic Procedures for Viral, Rickettsial and Chlamydial Infections, 5th Ed. American Public Health Association, Washington, DC, pp. 1–48.Google Scholar
  6. 6.
    Senne, D. A. (1998) Virus propagation in embryonating eggs. In: Swayne, D. E., Glisson, J. R., Jackwood, M. W., Pearson, J. E., and Reed, W. M. (eds.) A Laboratory Manual for Isolation and Identification of Avian Pathogens, 4th Ed., American Association of Avian Pathologists, Kennett Square, PA, pp. 235–240.Google Scholar
  7. 7.
    Cavanagh, D. (2003) Severe acute respiratory syndrome vaccine development: experiences of vaccination against avian infectious bronchitis virus Avian Pathol. 32, 567–582.PubMedCrossRefGoogle Scholar
  8. 8.
    Jordan, F. T. W., and Nassar, T. J. (1973) The combined influence of age of embryo, temperature and duration of incubation on the replication and yield of avian infectious bronchitis virus in the developing chick embryo. Avian Pathol. 2, 279–294.PubMedGoogle Scholar
  9. 9.
    Guy, J. S. (2000) Turkey coronavirus is more closely related to avian infectious bronchitis virus than to mammalian coronaviruses: a review Avian Pathol. 29, 207–212.PubMedCrossRefGoogle Scholar
  10. 10.
    Brake, J., Walsh T. J., Benton, C. E., Petitte, J. N, Meyerhof, R., and Penalva, G. (1997). Egg handling and storage Poult. Sci. 76, 144–151.PubMedGoogle Scholar
  11. 11.
    Jonassen, C. M., Kofstad, T., Larsen, I. L., Lovland, A., Handeland, K., Follestad, and A., Lillehaug, A. (2005) Molecular identification and characterization of novel coronaviruses infecting graylag geese (Anser anser), feral pigeons (Columba livia) and mallards (Anas platyrhynchos). J. Gen. Virol. 86, 1597–1607.PubMedCrossRefGoogle Scholar
  12. 12.
    Stephensen, C. B., Casebolt, D. B., and Gangopadhyay, N. N.(1999) Phylogenetic analysis of a highly conserved region of the polymerase gene from 11 coronaviruses and development of a consensus polymerase chain reaction assay Virus Res 60, 181–189.PubMedCrossRefGoogle Scholar
  13. 13.
    McNulty, M. S., Curran, W. L., Todd, D., and McFerran, J. B. (1979) Detection of viruses in avian faeces by direct electron microscopy Avian Pathol. 8, 239–247.PubMedCrossRefGoogle Scholar
  14. 14.
    Cavanagh, D., Mawditt, K., Welchman, D. B, Britton, P., and Gough, R.E. (2002) Coronaviruses from pheasants (Phasianus colchicus) are genetically closely related to coronaviruses of domestic fowl (infectious bronchitis virus) and turkeys. Avian Pathol. 31, 181–193.Google Scholar
  15. 15.
    Guy, J. S., Barnes, H. J., Smith, L. G., and Breslin, J. (1997) Antigenic characterization of a turkey coronavirus identified in poult enteritis and mortality syndrome-affected turkeys. Avian Dis. 41, 583–590.PubMedCrossRefGoogle Scholar
  16. 16.
    Cavanagh, D. (2005) Coronaviruses in poultry and other birds Avian Pathol. 23, 439–448.CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  • James S. Guy
    • 1
  1. 1.College of Veterinary MedicineNorth Carolina State UniversityRaleighUSA

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