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. 1995 Mar;33(3):684–689. doi: 10.1128/jcm.33.3.684-689.1995

Detection of feline coronavirus RNA in feces, tissues, and body fluids of naturally infected cats by reverse transcriptase PCR.

A A Herrewegh 1, R J de Groot 1, A Cepica 1, H F Egberink 1, M C Horzinek 1, P J Rottier 1
PMCID: PMC228014  PMID: 7751377

Abstract

A nested reverse transcriptase PCR (RT-nPCR) was developed for the detection of feline coronavirus (FCoV) RNA in the feces, tissues, and body fluids of infected cats. The RT-nPCR was targeted to the highly conserved 3'-untranslated region of the viral genome and will detect most, if not all, feline coronaviruses in the field. With the RT-nPCR, FCoV RNA was detected in plasma samples from experimentally infected cats as early as 2 days postinoculation. FCoV RNA was also detected in serum, plasma, or ascitic fluid samples from 14 of 18 cats (78%) with naturally occurring feline infectious peritonitis (FIP). The use of RT-PCR for FIP diagnosis is limited because of the occurrence of apparently healthy FCoV carriers. These asymptomatic cats shed the virus in the feces and, in a number of cases, also had detectable virus in the plasma. Because of the nature of FCoV infections, our RT-PCR assay with plasma or serum cannot be used to establish a definite diagnosis of FIP. However, this assay does provide a new means to identify asymptomatic FCoV carriers. As such, RT-nPCR will be of use to screen cats before their introduction into FCoV-free catteries. Moreover, this assay provides an important tool to study the epidemiology of FCoV.

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Selected References

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  1. Addie D. D., Jarrett J. O. Feline coronavirus antibodies in cats. Vet Rec. 1992 Aug 29;131(9):202–203. doi: 10.1136/vr.131.9.202-a. [DOI] [PubMed] [Google Scholar]
  2. Addie D. D., Jarrett O. A study of naturally occurring feline coronavirus infections in kittens. Vet Rec. 1992 Feb 15;130(7):133–137. doi: 10.1136/vr.130.7.133. [DOI] [PubMed] [Google Scholar]
  3. Barlough J. E. Serodiagnostic aids and management practice for feline retrovirus and coronavirus infections. Vet Clin North Am Small Anim Pract. 1984 Sep;14(5):955–969. doi: 10.1016/S0195-5616(84)50101-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barlough J. E., Stoddart C. A., Sorresso G. P., Jacobson R. H., Scott F. W. Experimental inoculation of cats with canine coronavirus and subsequent challenge with feline infectious peritonitis virus. Lab Anim Sci. 1984 Dec;34(6):592–597. [PubMed] [Google Scholar]
  5. Black J. W. Recovery and in vitro cultivation of a coronavirus from laboratory-induced cases of feline infectious peritonitis (FIP). Vet Med Small Anim Clin. 1980 May;75(5):811–814. [PubMed] [Google Scholar]
  6. Boom R., Sol C. J., Salimans M. M., Jansen C. L., Wertheim-van Dillen P. M., van der Noordaa J. Rapid and simple method for purification of nucleic acids. J Clin Microbiol. 1990 Mar;28(3):495–503. doi: 10.1128/jcm.28.3.495-503.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Boyle J. F., Pedersen N. C., Evermann J. F., McKeirnan A. J., Ott R. L., Black J. W. Plaque assay, polypeptide composition and immunochemistry of feline infectious peritonitis virus and feline enteric coronavirus isolates. Adv Exp Med Biol. 1984;173:133–147. doi: 10.1007/978-1-4615-9373-7_12. [DOI] [PubMed] [Google Scholar]
  8. Corapi W. V., Olsen C. W., Scott F. W. Monoclonal antibody analysis of neutralization and antibody-dependent enhancement of feline infectious peritonitis virus. J Virol. 1992 Nov;66(11):6695–6705. doi: 10.1128/jvi.66.11.6695-6705.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. De Groot R. J., Andeweg A. C., Horzinek M. C., Spaan W. J. Sequence analysis of the 3'-end of the feline coronavirus FIPV 79-1146 genome: comparison with the genome of porcine coronavirus TGEV reveals large insertions. Virology. 1988 Dec;167(2):370–376. doi: 10.1016/0042-6822(88)90097-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Evermann J. F., Baumgartener L., Ott R. L., Davis E. V., McKeirnan A. J. Characterization of a feline infectious peritonitis virus isolate. Vet Pathol. 1981 Mar;18(2):256–265. doi: 10.1177/030098588101800214. [DOI] [PubMed] [Google Scholar]
  11. Fiscus S. A., Teramoto Y. A. Antigenic comparison of feline coronavirus isolates: evidence for markedly different peplomer glycoproteins. J Virol. 1987 Aug;61(8):2607–2613. doi: 10.1128/jvi.61.8.2607-2613.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Goitsuka R., Hirota Y., Hasegawa A., Tomoda I. Release of interleukin 1 from peritoneal exudate cells of cats with feline infectious peritonitis. Nihon Juigaku Zasshi. 1987 Oct;49(5):811–818. doi: 10.1292/jvms1939.49.811. [DOI] [PubMed] [Google Scholar]
  13. Hohdatsu T., Okada S., Koyama H. Characterization of monoclonal antibodies against feline infectious peritonitis virus type II and antigenic relationship between feline, porcine, and canine coronaviruses. Arch Virol. 1991;117(1-2):85–95. doi: 10.1007/BF01310494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hohdatsu T., Sasamoto T., Okada S., Koyama H. Antigenic analysis of feline coronaviruses with monoclonal antibodies (MAbs): preparation of MAbs which discriminate between FIPV strain 79-1146 and FECV strain 79-1683. Vet Microbiol. 1991 Jun;28(1):13–24. doi: 10.1016/0378-1135(91)90096-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Horsburgh B. C., Brierley I., Brown T. D. Analysis of a 9.6 kb sequence from the 3' end of canine coronavirus genomic RNA. J Gen Virol. 1992 Nov;73(Pt 11):2849–2862. doi: 10.1099/0022-1317-73-11-2849. [DOI] [PubMed] [Google Scholar]
  16. McArdle F., Bennett M., Gaskell R. M., Tennant B., Kelly D. F., Gaskell C. J. Induction and enhancement of feline infectious peritonitis by canine coronavirus. Am J Vet Res. 1992 Sep;53(9):1500–1506. [PubMed] [Google Scholar]
  17. Montali R. J., Strandberg J. D. Extraperitoneal lesions in feline infectious peritonitis. Vet Pathol. 1972;9(2):109–121. doi: 10.1177/030098587200900204. [DOI] [PubMed] [Google Scholar]
  18. O'Reilly K. J., Fishman B., Hitchcock L. M. Feline infectious peritonitis: isolation of a coronavirus. Vet Rec. 1979 Apr 14;104(15):348–348. doi: 10.1136/vr.104.15.348. [DOI] [PubMed] [Google Scholar]
  19. Pedersen N. C., Black J. W., Boyle J. F., Evermann J. F., McKeirnan A. J., Ott R. L. Pathogenic differences between various feline coronavirus isolates. Adv Exp Med Biol. 1984;173:365–380. doi: 10.1007/978-1-4615-9373-7_36. [DOI] [PubMed] [Google Scholar]
  20. Pedersen N. C., Boyle J. F., Floyd K., Fudge A., Barker J. An enteric coronavirus infection of cats and its relationship to feline infectious peritonitis. Am J Vet Res. 1981 Mar;42(3):368–377. [PubMed] [Google Scholar]
  21. Pedersen N. C., Boyle J. F., Floyd K. Infection studies in kittens, using feline infectious peritonitis virus propagated in cell culture. Am J Vet Res. 1981 Mar;42(3):363–367. [PubMed] [Google Scholar]
  22. Pedersen N. C. Morphologic and physical characteristics of feline infectious peritonitis virus and its growth in autochthonous peritoneal cell cultures. Am J Vet Res. 1976 May;37(5):567–572. [PubMed] [Google Scholar]
  23. Pedersen N. C. Serologic studies of naturally occurring feline infectious peritonitis. Am J Vet Res. 1976 Dec;37(12):1449–1453. [PubMed] [Google Scholar]
  24. Pedersen N. C. Virologic and immunologic aspects of feline infectious peritonitis virus infection. Adv Exp Med Biol. 1987;218:529–550. doi: 10.1007/978-1-4684-1280-2_69. [DOI] [PubMed] [Google Scholar]
  25. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  26. Sparkes A. H., Gruffydd-Jones T. J., Harbour D. A. Feline coronavirus antibodies in UK cats. Vet Rec. 1992 Sep 5;131(10):223–224. doi: 10.1136/vr.131.10.223-a. [DOI] [PubMed] [Google Scholar]
  27. Sparkes A. H., Gruffydd-Jones T. J., Harbour D. A. Feline infectious peritonitis: a review of clinicopathological changes in 65 cases, and a critical assessment of their diagnostic value. Vet Rec. 1991 Sep 7;129(10):209–212. doi: 10.1136/vr.129.10.209. [DOI] [PubMed] [Google Scholar]
  28. Sparkes A. H., Gruffydd-Jones T. J., Howard P. E., Harbour D. A. Coronavirus serology in healthy pedigree cats. Vet Rec. 1992 Jul 11;131(2):35–36. doi: 10.1136/vr.131.2.35. [DOI] [PubMed] [Google Scholar]
  29. Stoddart C. A., Scott F. W. Intrinsic resistance of feline peritoneal macrophages to coronavirus infection correlates with in vivo virulence. J Virol. 1989 Jan;63(1):436–440. doi: 10.1128/jvi.63.1.436-440.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Vennema H., Rossen J. W., Wesseling J., Horzinek M. C., Rottier P. J. Genomic organization and expression of the 3' end of the canine and feline enteric coronaviruses. Virology. 1992 Nov;191(1):134–140. doi: 10.1016/0042-6822(92)90174-N. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ward J. M. Morphogenesis of a virus in cats with experimental feline infectious peritonitis. Virology. 1970 May;41(1):191–194. doi: 10.1016/0042-6822(70)90070-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weiss R. C., Scott F. W. Pathogenesis of feline infectious peritonitis: nature and development of viremia. Am J Vet Res. 1981 Mar;42(3):382–390. [PubMed] [Google Scholar]
  33. Witte K. H., Tuch K., Dubenkropp H., Walther C. Untersuchungen über die Antigenverwandtschaft der Viren der Felinen Infektiösen Peritonitis (FIP) und der Transmissiblen Gastroenteritis (TGE) des Schweines. Berl Munch Tierarztl Wochenschr. 1977 Oct 15;90(20):396–401. [PubMed] [Google Scholar]
  34. de Groot R. J., ter Haar R. J., Horzinek M. C., van der Zeijst B. A. Intracellular RNAs of the feline infectious peritonitis coronavirus strain 79-1146. J Gen Virol. 1987 Apr;68(Pt 4):995–1002. doi: 10.1099/0022-1317-68-4-995. [DOI] [PubMed] [Google Scholar]

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