Skip to main content
PMC home page
Elsevier - PMC COVID-19 Collection logoLink to Elsevier - PMC COVID-19 Collection
. 2002 Nov 12;21(3):263–273. doi: 10.1016/0378-1135(90)90037-V

Enzyme-linked immunosorbent assay for the detection of porcine epidemic diarrhea coronavirus antibodies in swine sera

Martin Hofmann 1, Robert Wyler 1,∗
PMCID: PMC7117443  PMID: 2154877

Abstract

An enzyme-linked immunosorbent assay (ELISA) for detecting serum antibodies to the porcine epidemic diarrhea coronavirus (PEDV) was established by using cell culture-grown PEDV as antigen for coating. Ultracentrifugation through 20 and 45% (w/w) sucrose cushion proved to be the best antigen purification method. Examination of 1024 swine sera showed a high specificity and a greater sensitivity of the ELISA, when compared with indirect immunofluorescence. Reference sera with high antibody titers to PEDV originated from two pigs experimentally infected with PEDV. Three different antigen purification methods and the advantages of the ELISA compared with an immunofluorescence test are discussed.

References

  1. Bereiter M., Hasler J., Keller H. Transmissible Gastroenteritis (TGE) in der Schweiz: Antikörperpersistenz nach Infektion und seroepidemiologische Untersuchungen zur Bedeuting des TGE-Virus als Durchfallerreger. Schweiz. Arch. Tierheilkd. 1988;130:237–248. [PubMed] [Google Scholar]
  2. Callebaut P., Debouck P., Pensaert M. Enzyme-linked immunosorbent assay for the detection of the coronavirus-like agent and its antibodies in pigs with porcine epidemic diarrhea. Vet. Microbiol. 1982;7:295–306. doi: 10.1016/0378-1135(82)90009-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Debouck P., Pensaert M. Experimental infection of pigs with a new porcine enteric coronavirus, CV777. Am. J. Vet. Res. 1980;41:219–223. [PubMed] [Google Scholar]
  4. Debouck P., Pensaert M., Coussement W. The pathogenesis of an enteric infection in pigs, experimentally induced by the coronavirus-like agent, CV 777. Vet. Microbiol. 1981;6:157–165. [Google Scholar]
  5. Gardas A., Lewartowska A. Coating of proteins to polystyrene ELISA plates in the presence of detergents. J. Immunol. Methods. 1988;106:251–255. doi: 10.1016/0022-1759(88)90205-0. [DOI] [PubMed] [Google Scholar]
  6. Hofmann M., Wyler R. Serologische Untersuchung über das Vorkommen der Epizootischen Virusdiarrhoe der Schweine (EVD) in der Schweiz. Schweiz. Arch. Tierheilkd. 1987;129:437–442. [PubMed] [Google Scholar]
  7. Hofmann M., Wyler R. Propagation of the virus of porcine epidemic diarrhea in cell culture. J. Clin. Microbiol. 1988;26:2235–2239. doi: 10.1128/jcm.26.11.2235-2239.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jimenez G., Correa I., Melgosa M.P., Bullido M.J., Enjuanes L. Critical epitopes in transmissible gastroenteritis virus neutralization. J. Virol. 1986;60:131–139. doi: 10.1128/jvi.60.1.131-139.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kenny G.E., Dunsmoor C.L. Principles, problems, and strategies in the use of antigenic mixtures for the enzyme-linked immunosorbent assay. J. Clin. Microbiol. 1983;17:655–665. doi: 10.1128/jcm.17.4.655-665.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Pensaert M.B., Debouck P. A new coronavirus-like agent associated with diarrhea in swine. Arch. Virol. 1978;58:243–247. doi: 10.1007/BF01317606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Prager D., Witte K.H. Die serologische Diagnose der Epizootischen Virusdiarrhoe (EVD) des Schweines mit Hilfe der indirekten Immunfluoreszenztechnik (IIFT). I. Einfluss verschiedener Parameter auf den Antikörpertiter. Tieraerzt. Umsch. 1981;36:404–414. [Google Scholar]
  12. Prager D., Witte K.H. Die serologische Diagnose der Epizootischen Virusdiarrhoe (EVD) des Schweines mit Hilfe der indirekten Immunfluoreszenztechnik (IIFT). II. Antikörper-Antwort nach experimenteller Infektion. Tieraerztl. Umsch. 1981;36:477–480. [Google Scholar]
  13. Siddell S.G., Anderson R., Cavanagh D., Fujiwara K., Klenk H.D., Macnaughton M.R., Pensaert M., Stohlman S.A., Sturman L., Vanderzeijst B.A.M. Coronaviridae. Intervirology. 1983;20:181–189. doi: 10.1159/000149390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sturman L.S., Holmes K.V. The molecular biology of coronaviruses. In: Lauffer M.A., Maramorosch K., editors. Vol. 28. Academic Press; New York: 1983. pp. 35–112. (Advances in Virus Research). [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sturman L.S., Holmes K.V., Behnke J. Isolation of coronavirus envelope glycoproteins and interaction with the viral nucleocapsid. J. Virol. 1980;33:449–462. doi: 10.1128/jvi.33.1.449-462.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Witte K.H., Prager D. Der Nachweis von Antikörpern gegen das Virus der Epizootischen Virusdiarrhoe (EVD) des Schweines mit dem Immunfluoreszenzblockadetest (IFBT) Tieraerztl. Umsch. 1987;42:817–820. [Google Scholar]
  17. Witte K.H., Prager D., Ernst H., Nienhoff H. Die Epizootische Virusdiarrhoe (EVD) Tieraerztl. Umsch. 1981;36:235–250. [Google Scholar]

Articles from Veterinary Microbiology are provided here courtesy of Elsevier

RESOURCES