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. 1988 Nov;26(11):2235–2239. doi: 10.1128/jcm.26.11.2235-2239.1988

Propagation of the virus of porcine epidemic diarrhea in cell culture.

M Hofmann 1, R Wyler 1
PMCID: PMC266866  PMID: 2853174

Abstract

Porcine epidemic diarrhea virus (PEDV) was adapted to serial propagation in Vero cell cultures by adding trypsin to the medium. PEDV-infected cells showed a distinct cytoplasmic fluorescence when examined by a fluorescent-antibody-staining technique. Cytopathic effects, such as vacuolation, formation of syncytia, and fusion of cells, were detected even at passage 1 of the PEDV in Vero cells. Once adapted, the virus induced numerous syncytia containing over 100 nuclei. From virus passage 5 on, all cells forming the monolayer were fused and totally destroyed within 24 h after inoculation. Cell culture-grown PEDV had typical coronavirus morphology when viewed by electron microscopy. Attempts to propagate PEDV in several primary and secondary fetal porcine cell cultures in the presence or absence of trypsin were unsuccessful.

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Selected References

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

  1. Chasey D., Cartwright S. F. Virus-like particles associated with porcine epidemic diarrhoea. Res Vet Sci. 1978 Sep;25(2):255–256. doi: 10.1016/S0034-5288(18)32994-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cyr-Coats K. S., Storz J. Bovine coronavirus-induced cytopathic expression and plaque formation: host cell and virus strain determine trypsin dependence. Zentralbl Veterinarmed B. 1988 Jan;35(1):48–56. doi: 10.1111/j.1439-0450.1988.tb00465.x. [DOI] [PubMed] [Google Scholar]
  3. Dea S., Roy R. S., Begin M. E. Bovine coronavirus isolation and cultivation in continuous cell lines. Am J Vet Res. 1980 Jan;41(1):30–38. [PubMed] [Google Scholar]
  4. Dea S., Roy R. S., Begin M. E. Physicochemical and biological properties of neonatal calf diarrhea coronaviruses isolated in Quebec and comparison with the Nebraska calf coronavirus. Am J Vet Res. 1980 Jan;41(1):23–29. [PubMed] [Google Scholar]
  5. Dea S., Vaillancourt J., Elazhary Y., Martineau G. P. An outbreak of diarrhea in piglets caused by a coronavirus antigenically distinct from transmissible gastroenteritis virus. Can Vet J. 1985 Mar;26(3):108–111. [PMC free article] [PubMed] [Google Scholar]
  6. Debouck P., Pensaert M. Experimental infection of pigs with a new porcine enteric coronavirus, CV 777. Am J Vet Res. 1980 Feb;41(2):219–223. [PubMed] [Google Scholar]
  7. Flynn W. T., Saif L. J. Serial propagation of porcine enteric calicivirus-like virus in primary porcine kidney cell cultures. J Clin Microbiol. 1988 Feb;26(2):206–212. doi: 10.1128/jcm.26.2.206-212.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Frana M. F., Behnke J. N., Sturman L. S., Holmes K. V. Proteolytic cleavage of the E2 glycoprotein of murine coronavirus: host-dependent differences in proteolytic cleavage and cell fusion. J Virol. 1985 Dec;56(3):912–920. doi: 10.1128/jvi.56.3.912-920.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gerna G., Cattaneo E., Cereda P. M., Revelo M. G., Achilli G. Human coronavirus OC43 serum inhibitor and neutralizing antibody by a new plaque-reduction assay. Proc Soc Exp Biol Med. 1980 Mar;163(3):360–366. doi: 10.3181/00379727-163-40778. [DOI] [PubMed] [Google Scholar]
  10. Hess R. G., Bollwahn W., Pospischil A., Heinritzi K., Bachmann P. A. Neue Aspekte der Virusätiologie bei Durchfallerkrankungen des Schweines: Vorkommen von Infektionen mit dem Epizootischen Virusdiarrhoe- (EVD-) virus. Berl Munch Tierarztl Wochenschr. 1980 Nov;93(22):445–449. [PubMed] [Google Scholar]
  11. Hirano N., Ono K., Sada Y., Inoue A., Murakami T., Takamaru H. Replication of rat coronavirus in a rat cell line, LBC. Brief report. Arch Virol. 1985;85(3-4):301–304. doi: 10.1007/BF01314238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hirano N., Sada Y., Tuchiya K., Ono K., Murakami T. Plaque assay of bovine coronavirus in BEK-1 cells. Nihon Juigaku Zasshi. 1985 Aug;47(4):679–681. doi: 10.1292/jvms1939.47.679. [DOI] [PubMed] [Google Scholar]
  13. Hooper B. E., Haelterman E. O. Growth of transmissible gastroenteritis virus in young pigs. Am J Vet Res. 1966 Jan;27(116):286–291. [PubMed] [Google Scholar]
  14. Horváth I., Mocsári E. Ultrastructural changes in the small intestinal epithelium of suckling pigs affected with a transmissible gastroenteritis (TGE)-like disease. Arch Virol. 1981;68(2):103–113. doi: 10.1007/BF01314440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Komaniwa H., Makabe T., Fukusho A., Shimizu Y. Isolation of transmissible gastroenteritis virus from feces of diarrheic pigs in roller culture of CPK cells in the presence of trypsin. Nihon Juigaku Zasshi. 1986 Dec;48(6):1245–1248. doi: 10.1292/jvms1939.48.1245. [DOI] [PubMed] [Google Scholar]
  16. Lee T. W., Kurtz J. B. Serial propagation of astrovirus in tissue culture with the aid of trypsin. J Gen Virol. 1981 Dec;57(Pt 2):421–424. doi: 10.1099/0022-1317-57-2-421. [DOI] [PubMed] [Google Scholar]
  17. MADIN S. H., ANDRIESE P. C., DARBY N. B. The in vitro cultivation of tissues of domestic and laboratory animals. Am J Vet Res. 1957 Oct;18(69):932–941. [PubMed] [Google Scholar]
  18. Pensaert M. B., Debouck P., Reynolds D. J. An immunoelectron microscopic and immunofluorescent study on the antigenic relationship between the coronavirus-like agent, CV 777, and several coronaviruses. Arch Virol. 1981;68(1):45–52. doi: 10.1007/BF01315166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pensaert M. B., Haelterman E. O., Burnstein T. Diagnosis of transmissible gastroenteritis in pigs by means of immunofluorescence. Can J Comp Med. 1968 Oct;32(4):555–561. [PMC free article] [PubMed] [Google Scholar]
  20. Pensaert M. B., de Bouck P. A new coronavirus-like particle associated with diarrhea in swine. Arch Virol. 1978;58(3):243–247. doi: 10.1007/BF01317606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Robb J. A., Bond C. W. Pathogenic murine coronaviruses. I. Characterization of biological behavior in vitro and virus-specific intracellular RNA of strongly neurotropic JHMV and weakly neurotropic A59V viruses. Virology. 1979 Apr 30;94(2):352–370. doi: 10.1016/0042-6822(79)90467-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Siddell S., Wege H., Ter Meulen V. The biology of coronaviruses. J Gen Virol. 1983 Apr;64(Pt 4):761–776. doi: 10.1099/0022-1317-64-4-761. [DOI] [PubMed] [Google Scholar]
  23. Storz J., Rott R., Kaluza G. Enhancement of plaque formation and cell fusion of an enteropathogenic coronavirus by trypsin treatment. Infect Immun. 1981 Mar;31(3):1214–1222. doi: 10.1128/iai.31.3.1214-1222.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sturman L. S., Holmes K. V. The molecular biology of coronaviruses. Adv Virus Res. 1983;28:35–112. doi: 10.1016/S0065-3527(08)60721-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sturman L. S., Ricard C. S., Holmes K. V. Proteolytic cleavage of the E2 glycoprotein of murine coronavirus: activation of cell-fusing activity of virions by trypsin and separation of two different 90K cleavage fragments. J Virol. 1985 Dec;56(3):904–911. doi: 10.1128/jvi.56.3.904-911.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Toth T. E. Trypsin-enhanced replication of neonatal calf diarrhea coronavirus in bovine embryonic lung cells. Am J Vet Res. 1982 Jun;43(6):967–972. [PubMed] [Google Scholar]
  27. Turgeon D. C., Morin M., Jolette J., Higgins R., Marsolais G., DiFranco E. Coronavirus-like particles associated with diarrhea in baby pigs in Quebec. Can Vet J. 1980 Mar;21(3):100–xxiii. [PMC free article] [PubMed] [Google Scholar]
  28. Tyrrell D. A., Alexander D. J., Almeida J. D., Cunningham C. H., Easterday B. C., Garwes D. J., Hierholzer J. C., Kapikian A., Macnaughton M. R., McIntosh K. Coronaviridae: second report. Intervirology. 1978;10(6):321–328. doi: 10.1159/000148996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Vautherot J. F. Plaque assay for titration of bovine enteric coronavirus. J Gen Virol. 1981 Oct;56(Pt 2):451–455. doi: 10.1099/0022-1317-56-2-451. [DOI] [PubMed] [Google Scholar]
  30. Witte K. H., Easterday B. C. Isolation and propagation of the virus of transmissible gastroenteritis of pigs in various pig cell cultures. Arch Gesamte Virusforsch. 1967;20(3):327–350. doi: 10.1007/BF01241952. [DOI] [PubMed] [Google Scholar]
  31. Witte K. H. Isolation of the virus of transmissible gastroenteritis (TGE) from naturally infected piglets in cell culture. Zentralbl Veterinarmed B. 1971 Dec;18(10):770–778. doi: 10.1111/j.1439-0450.1971.tb01654.x. [DOI] [PubMed] [Google Scholar]
  32. Wood E. N. An apparently new syndrome of porcine epidemic diarrhoea. Vet Rec. 1977 Mar 19;100(12):243–244. doi: 10.1136/vr.100.12.243. [DOI] [PubMed] [Google Scholar]
  33. Wood E. N. Transmissible gastroenteritis and epidemic diarrhoea of pigs. Br Vet J. 1979 Jul-Aug;135(4):305–314. doi: 10.1016/s0007-1935(17)32831-2. [DOI] [PubMed] [Google Scholar]
  34. Wyatt R. G., James W. D., Bohl E. H., Theil K. W., Saif L. J., Kalica A. R., Greenberg H. B., Kapikian A. Z., Chanock R. M. Human rotavirus type 2: cultivation in vitro. Science. 1980 Jan 11;207(4427):189–191. doi: 10.1126/science.6243190. [DOI] [PubMed] [Google Scholar]

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