Skip to main content

Isolation and Propagation of Coronaviruses in Embryonated Eggs

  • Protocol
  • First Online:
Coronaviruses

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2203))

Abstract

The embryonated egg is a complex structure comprised of an embryo and its supporting membranes (chorioallantoic, amniotic, and yolk). The developing embryo and its membranes provide a diversity of cell types that allow for the 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 avian coronaviruses within the genus Gammacoronavirus, infectious bronchitis virus (IBV) and turkey coronavirus (TCoV). IBV replicates well in the embryonated chicken egg, regardless of inoculation route; however, the allantoic route is favored as the virus replicates well in epithelium lining the chorioallantoic membrane, with high virus titers found in these membranes and associated allantoic fluids. TCoV replicates only in epithelium lining the embryo 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 detection, propagation, and characterization of other, novel coronaviruses.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 249.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Adams NR, Hofstad MS (1970) Isolation of transmissible enteritis agent of turkeys in avian embryos. Avian Dis 15:426–433

    Article  Google Scholar 

  2. Cavanagh D, Naqi SA (2003) Infectious bronchitis. In: Saif YM, Barnes HJ, Fadly A, Glisson JR, McDougald LR, Swayne DE (eds) Diseases of poultry, 11th edn. Iowa State University Press, Ames, Iowa, pp 101–120

    Google Scholar 

  3. Guy JS (2013) Turkey coronavirus enteritis. In: Swayne DE, Glisson JR, McDougald LR (eds) Diseases of poultry, 13th edn. Ames, Iowa, Wiley-Blackwell, pp 376–381

    Google Scholar 

  4. Gough RE, Cox WJ, Winkler CE, Sharp MW, Spackman D (1996) Isolation and identification of infectious bronchitis virus from pheasants. Vet Rec 138:208–209

    Article  CAS  Google Scholar 

  5. Liu S, Chen J, Kong X, Shao Y, Han Z, Feng L, Cai X, Gu S, Liu M (2005) Isolation of infectious bronchitis coronavirus from domestic peafowl (Pavo cristatus) and teal (Anas). J Gen Virol 86:719–725

    Article  CAS  Google Scholar 

  6. Qian DH, Zhu GJ, Wu LZ, Hua GX (2006) Isolation and characterization of a coronavirus from pigeons with pancreatitis. Am J Vet Res 67:1575–1579

    Article  CAS  Google Scholar 

  7. Hawkes RA (1979) General principals underlying laboratory diagnosis of viral infections. In: Lennette EH, Schmidt NJ (eds) Diagnostic procedures for viral, rickettsial and chlamydial infections, 5th edn. American Public Health Association, Washington, DC, pp 1–48

    Google Scholar 

  8. Senne DA (2008) Virus propagation in embryonating eggs. In: Dufour-Zavala L, Swayne DE, Glisson JR, Pearson JE, Reed WM, Jackwood MW, Woolcock PR (eds) A laboratory manual for isolation and identification of avian pathogens, 5th edn. American Association of Avian Pathologists, Jacksonville, FL, pp 204–208

    Google Scholar 

  9. Cavanagh D (2003) Severe acute respiratory syndrome vaccine development: experiences of vaccination against avian infectious bronchitis virus. Avian Pathol 32:567–582

    Article  CAS  Google Scholar 

  10. Jordan FTW, Nassar TJ (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

    Article  CAS  Google Scholar 

  11. Guy JS (2000) Turkey coronavirus is more closely related to avian infectious bronchitis virus that to mammalian coronaviruses: a review. Avian Pathol 29:207–212

    Article  CAS  Google Scholar 

  12. Brake J, Walsh TJ, Benton CE, Petitte JN, Meyerhof R, Penalva (1997) Egg handling and storage. Poult Sci 76:144–151

    Article  CAS  Google Scholar 

  13. Jonassen CM, Kofstad T, Larsen IL, Lovland A, Handeland K, Follestad A, Lillehaug A (2005) Molecular identification and characterization of novel coronaviruses infecting graylag geese (Anser anser), feral pigeons (Columag livia) and mallards (Anas platyrhynchos). J Gen Virol 86:1597–1607

    Article  CAS  Google Scholar 

  14. Stephensen CB, Casebolt DB, Gangopadhyay NN (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

    Article  CAS  Google Scholar 

  15. McNulty MS, Curran WL, Todd D, McFerran JB (1979) Detection of viruses in avian faeces by direct electron microscopy. Avian Pathol 8:239–247

    Article  CAS  Google Scholar 

  16. Cavanagh D, Mawditt K, Welchman DB, Britton P, Gough RE (2002) Coronaviruses from pheasants (Phasianus colchicus) are genetically closely related to coronaviruses of domestic fowl (infectious bronchitis virus) and turkeys. Avian Pathol 31:81–93

    Article  CAS  Google Scholar 

  17. Guy JS, Barnes HJ, Smith LG, Breslin J (1997) Antigenic characterization of a Turkey coronavirus identified in poult enteritis and mortality syndrome-affected turkeys. Avian Dis 41:583–590

    Article  CAS  Google Scholar 

  18. Cavanagh D (2005) Coronaviruses in poultry and other birds. Avian Pathol 34:439–448

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to James S. Guy .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Guy, J.S. (2020). Isolation and Propagation of Coronaviruses in Embryonated Eggs. In: Maier, H., Bickerton, E. (eds) Coronaviruses. Methods in Molecular Biology, vol 2203. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0900-2_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0900-2_9

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0899-9

  • Online ISBN: 978-1-0716-0900-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics