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. 1993 Oct;57(4):300–304.

Immunohistochemical detection of porcine reproductive and respiratory syndrome virus using colloidal gold.

R Magar 1, R Larochelle 1, Y Robinson 1, C Dubuc 1
PMCID: PMC1263644  PMID: 8269370

Abstract

Two cytopathic agents were isolated on porcine alveolar macrophages following inoculation with homogenates of lung tissues from pigs showing respiratory problems. These isolates were identified as porcine reproductive and respiratory syndrome (PRRS) virus isolates by indirect immunofluorescence using a PRRS virus (PRRSV) specific monoclonal antibody (MAb) and were designated as LHVA-92-1 and LHVA-92-2. Immunogold electron microscopy using a porcine PRRS positive serum pool and protein A-gold resulted in an intense labelling of aggregates of viral particles. Dark specific cytoplasmic staining of porcine alveolar macrophages infected with both virus isolates could be observed by immunogold silver staining (IGSS) using the specific MAb. This method proved effective in detecting PRRSV antigens in several ethanol-fixed tissues of piglets intranasally inoculated with the supernatants of macrophages infected with each isolate. Immunogold silver staining was also successfully used for the detection of PRRSV antigens on sections of formalin-fixed paraffin-embedded lung tissues and on frozen sections of lungs. The present results indicate that colloidal gold may be useful for the identification and immunohistochemical detection of PRRSV in tissues.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bouillant A. M., Dulac G. C., Willis N., Girard A., Greig A. S., Boulanger P. Viral susceptibility of a cell line derived from the pig oviduct. Can J Comp Med. 1975 Oct;39(4):450–456. [PMC free article] [PubMed] [Google Scholar]
  2. Collins J. E., Benfield D. A., Christianson W. T., Harris L., Hennings J. C., Shaw D. P., Goyal S. M., McCullough S., Morrison R. B., Joo H. S. Isolation of swine infertility and respiratory syndrome virus (isolate ATCC VR-2332) in North America and experimental reproduction of the disease in gnotobiotic pigs. J Vet Diagn Invest. 1992 Apr;4(2):117–126. doi: 10.1177/104063879200400201. [DOI] [PubMed] [Google Scholar]
  3. Conzelmann K. K., Visser N., Van Woensel P., Thiel H. J. Molecular characterization of porcine reproductive and respiratory syndrome virus, a member of the arterivirus group. Virology. 1993 Mar;193(1):329–339. doi: 10.1006/viro.1993.1129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dea S., Bilodeau R., Sauvageau R., Montpetit C., Martineau G. P. Antigenic variant of swine influenza virus causing proliferative and necrotizing pneumonia in pigs. J Vet Diagn Invest. 1992 Oct;4(4):380–392. doi: 10.1177/104063879200400403. [DOI] [PubMed] [Google Scholar]
  5. Ghitescu L., Bendayan M. Immunolabeling efficiency of protein A-gold complexes. J Histochem Cytochem. 1990 Nov;38(11):1523–1530. doi: 10.1177/38.11.2212613. [DOI] [PubMed] [Google Scholar]
  6. Girard C., Morin M., el Azhary Y. Experimentally induced porcine proliferative and necrotising pneumonia with an influenza A virus. Vet Rec. 1992 Mar 7;130(10):206–207. doi: 10.1136/vr.130.10.206. [DOI] [PubMed] [Google Scholar]
  7. Hopley J. F., Doane F. W. Development of a sensitive protein A-gold immunoelectron microscopy method for detecting viral antigens in fluid specimens. J Virol Methods. 1985 Oct;12(1-2):135–147. doi: 10.1016/0166-0934(85)90014-x. [DOI] [PubMed] [Google Scholar]
  8. Larochelle R., Magar R. The application of immunogold silver staining (IGSS) for the detection of transmissible gastroenteritis virus in fixed tissues. J Vet Diagn Invest. 1993 Jan;5(1):16–20. doi: 10.1177/104063879300500105. [DOI] [PubMed] [Google Scholar]
  9. Magar R., Larochelle R. Immunohistochemical detection of porcine rotavirus using immunogold silver staining (IGSS). J Vet Diagn Invest. 1992 Jan;4(1):3–7. doi: 10.1177/104063879200400102. [DOI] [PubMed] [Google Scholar]
  10. McQuillin J., Madeley C. R., Kendal A. P. Monoclonal antibodies for the rapid diagnosis of influenza A and B virus infections by immunofluorescence. Lancet. 1985 Oct 26;2(8461):911–914. doi: 10.1016/s0140-6736(85)90849-9. [DOI] [PubMed] [Google Scholar]
  11. Meulenberg J. J., Hulst M. M., de Meijer E. J., Moonen P. L., den Besten A., de Kluyver E. P., Wensvoort G., Moormann R. J. Lelystad virus, the causative agent of porcine epidemic abortion and respiratory syndrome (PEARS), is related to LDV and EAV. Virology. 1993 Jan;192(1):62–72. doi: 10.1006/viro.1993.1008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Morin M., Girard C., Elazhary Y., Fajardo R., Drolet R., Lagacé A. Severe proliferative and necrotizing pneumonia in pigs: A newly recognized disease. Can Vet J. 1990 Dec;31(12):837–839. [PMC free article] [PubMed] [Google Scholar]
  13. Pol J. M., van Dijk J. E., Wensvoort G., Terpstra C. Pathological, ultrastructural, and immunohistochemical changes caused by Lelystad virus in experimentally induced infections of mystery swine disease (synonym: porcine epidemic abortion and respiratory syndrome (PEARS)). Vet Q. 1991 Jul;13(3):137–143. doi: 10.1080/01652176.1991.9694298. [DOI] [PubMed] [Google Scholar]
  14. Terpstra C., Wensvoort G., Pol J. M. Experimental reproduction of porcine epidemic abortion and respiratory syndrome (mystery swine disease) by infection with Lelystad virus: Koch's postulates fulfilled. Vet Q. 1991 Jul;13(3):131–136. doi: 10.1080/01652176.1991.9694297. [DOI] [PubMed] [Google Scholar]
  15. Welch S. K., Saif L. J. Monoclonal antibodies to a virulent strain of transmissible gastroenteritis virus: comparison of reactivity with virulent and attenuated virus. Arch Virol. 1988;101(3-4):221–235. doi: 10.1007/BF01311003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Wensvoort G., Terpstra C., Pol J. M., ter Laak E. A., Bloemraad M., de Kluyver E. P., Kragten C., van Buiten L., den Besten A., Wagenaar F. Mystery swine disease in The Netherlands: the isolation of Lelystad virus. Vet Q. 1991 Jul;13(3):121–130. doi: 10.1080/01652176.1991.9694296. [DOI] [PubMed] [Google Scholar]
  17. Yoon I. J., Joo H. S., Christianson W. T., Kim H. S., Collins J. E., Morrison R. B., Dial G. D. An indirect fluorescent antibody test for the detection of antibody to swine infertility and respiratory syndrome virus in swine sera. J Vet Diagn Invest. 1992 Apr;4(2):144–147. doi: 10.1177/104063879200400205. [DOI] [PubMed] [Google Scholar]

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