Skip to main content
  • Published:

In Situ Hybridization for the Detection of Infectious Laryngotracheitis Virus in Sections of Trachea from Experimentally Infected Chickens

Abstract

An in situ hybridization procedure for the detection of infectious laryngotracheitis virus (ILTV) in experimentally infected chickens is described. Formalin-fixed, paraffin-embedded sections of trachea, taken from chickens on days 3–10 post-inoculation (p.i.) with ILTV were hybridized with a mixture of 2 biotinylated, polymerase chain reaction-generated DNA fragments. The fragments correspond to sequences of the ILTV glycoprotein C and thymidine kinase genes. In situ hybridization was seen in 7 out of 7 chickens examined on day 3 p.i., 2 out of 2 examined on day 4 p.i. and 3 out of 3 examined on day 5 p.i. No hybridization was observed in 3 out of 3 chickens examined on day 10 p.i. ILTV nucleic acid was detected in nuclei of degenerated tracheal epithelial cells and in intranuclear inclusion bodies of syncytia.

References

  • Abbas F, Andreasen Jr. JR: Comparison of diagnostic tests for infectious laryngotracheitis. Avian Diseases, 1996, 40, 290–295.

    Article  CAS  Google Scholar 

  • Alexander DJ, Gough RE: Virus diseases of the respiratory organs. World situation and recent developments. Proceedings, Xlth International Congress of the World Veterinary Poultry Association, 18-22 August, 1997, Budapest, Hungary.

  • Anonymous: The 1993 Annual Report from the Danish National Veterinary Laboratory and State Veterinary Institute for Virus Research.

  • Bagust TJ, Calnek BW, Fahey KJ: Gallid-1 herpesvirus infection in the chicken. 3. Reinvestigation of the pathogenesis of infectious laryngotracheitis in acute and early post-acute respiratory disease. Avian Diseases, 1986, 30, 179–190.

    CAS  PubMed  Google Scholar 

  • Bagust TJ, Guy JS: Laryngotracheitis. In: Calnek BW, Barnes HJ, Beard CW, McDougald LR, Saif YM (eds.): Diseases of poultry. 10th Edn. Iowa State University Press, Ames, 1997, 527–539.

    Google Scholar 

  • Engström BE, Renström LHM, Wahlström H: Transmission of infectious laryngo-tracheitis in backyard chicken in Sweden. Abstract, Xlth International Congress of the World Veterinary Poultry Association, 18-22 August, 1997, Budapest, Hungary.

    Google Scholar 

  • GuyJS, Barnes HJ, Smith LG: Rapid diagnosis of infectious laryngotracheitis using a monoclonal antibody-based immunoperoxidase procedure. Avian Pathology, 1992, 21, 77–86.

    Article  Google Scholar 

  • Hughes CS, Jones RC: Comparison of cultural methods for primary isolation of infectious laryngotracheitis virus from field material. Avian Pathology, 1988, 77, 295–303.

    Article  Google Scholar 

  • Ide PR: Sensitivity and specificity of the fluorescent antibody technique for detection of infectious laryngotracheitis virus. Canadian Journal of Comparative Medicine, 1978, 42, 54–62.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Keam L, York J J, Sheppard M, Fahey KJ: Detection of infectious laryngotracheitis virus in chickens using a non-radioactive DNA probe. Avian Diseases, 1991, 35, 257–262.

    Article  CAS  Google Scholar 

  • Key DW, Gough BC, Derbyshire JB, Nagy E: Development and evaluation of a non-isotopically labelled DNA probe for the diagnosis of infectious laryngotracheitis. Avian Diseases, 1994, 38, 467–474.

    Article  CAS  Google Scholar 

  • Lan HY, Mu W, Ng YY, Nikolic-Paterson DJ, Atkins RC: A simple, reliable, and sensitive method for nonradioactive in situ hybridization: Use of microwave heating to improve hybridization efficiency and preserve tissue morphology. The Journal of Histochemistry and Cytochemistry, 1996, 44, 281–287.

    Article  CAS  Google Scholar 

  • Nielsen OL, Jorgensen PH, Bisgaard M, Alexandersen S: In situ hybridization for the detection of chicken anaemia virus in experimentally-induced infection and field outbreaks. Avian Pathology, 1995, 24, 149–155.

    Article  CAS  Google Scholar 

  • Riddell C: Avian Histopathology. 1st Edn. American Association of Avian Pathologists, University of Pennsylvania, 1987.

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T: Molecular cloning: A laboratory manual. 2nd Edn, Vol. 3. Cold Spring Harbor Laboratory Press, New York, 1989.

  • Shirley MW, Kemp DJ, Sheppard M, Fahey KJ: Detection of DNA from infectious laryngotracheitis virus by colourimetric analysis of polymerase chain reactions. Journal of Virological Methods, 1990, 30, 251–260.

    Article  CAS  Google Scholar 

  • Van Kämmen A, Spradbrow PB: Rapid diagnosis of some avian virus diseases. Avian Diseases, 1976, 20, 748–751.

    Article  Google Scholar 

  • Wilks CR, Kogan VG: An immunofluorescence diagnostic test for avian infectious laryngotracheitis. Australian Veterinary Journal, 1979, 55, 385–388.

    Article  CAS  Google Scholar 

  • Williams RA, Savage CE, Jones RC: A comparison of direct electron microscopy, virus isolation and a DNA amplification method for the detection of avian infectious laryngotracheitis virus in field material. Avian Pathology, 1994, 23, 709–720.

    Article  CAS  Google Scholar 

  • York J J, Fahey KJ: Diagnosis of infectious laryngotracheitis using a monoclonal antibody ELISA. Avian Pathology, 1988, 17, 173–182.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to O. L. Nielsen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nielsen, O.L., Handberg, K.J. & Jørgensen, P.H. In Situ Hybridization for the Detection of Infectious Laryngotracheitis Virus in Sections of Trachea from Experimentally Infected Chickens. Acta Vet Scand 39, 415–421 (1998). https://doi.org/10.1186/BF03547767

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1186/BF03547767