Abstract
An RT-PCR method was developed that amplified genetic material from the 5′ end of the S protein gene of both transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV), but discriminated between the two by the size of the product generated. A number of restriction endonuclease enzymes were assessed for recognition of the amplicons so produced. The assay was shown to detect viral RNA from all of the 26 different TGEV and PRCV isolates examined, covering a period from 1946 to 1996. Detection of TGEV in clinical specimens was possible using a spin column method to extract RNA and sensitivity was compared to virus isolation and antigen detection ELISA. The method could provide a means of confirming positive results from immunological screening tests such as FAT and ELISA, reducing the need for virus isolation and convalescent serology.
Keywords: Gastroenteritis virus, Respiratory coronavirus, Transmissible gastroenteritis
References
- Bae I., Jackwood D.J., Benfield D.A., Saif L.J., Wesley R.D., Hill H. Differentiation of transmissible gastroenteritis virus from porcine respiratory coronavirus and other antigenically related coronaviruses by using cDNA probes specific for the 5′ region of the S glycoprotein gene. J. Clin. Microbiol. 1991;29:215–218. doi: 10.1128/jcm.29.1.215-218.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bernard S., Lantier L., Laude H., Aynaud J.M. Detection of transmissible gastroenteritis coronavirus antigens by a sandwich enzyme-linked immunosorbent assay technique. Amer. J. Vet. Res. 1986;47:2441–2444. [PubMed] [Google Scholar]
- Callebaut P., Correa I., Pensaert M., Jimenez G., Enjuanes L. Antigenic differentiation between transmissible gastroenteritis of swine and a related porcine respiratory coronavirus. J. Gen. Virol. 1988;69:1725–1730. doi: 10.1099/0022-1317-69-7-1725. [DOI] [PubMed] [Google Scholar]
- Cartwright S.F., Harris H.M., Blandford T.B., Fincham I., Gitter M. A cytopathic virus causing a transmissible gastroenteritis in swine. I. Isolation and properties. J. Comp. Path. 1965;75:387–396. doi: 10.1016/0021-9975(65)90019-8. [DOI] [PubMed] [Google Scholar]
- Garwes D.J., Stewart F., Elleman C.J. Identification of epitopes of immunological importance on the peplomer of porcine transmissible gastroenteritis virus. In: Lai M.M.C., Stohlman S.A., editors. Coronaviruses, Advances in Experimental Medicine and Biology. 1987. pp. 509–515. (218) [DOI] [PubMed] [Google Scholar]
- Garwes D.J., Stewart F., Cartwright S.F., Brown I. Differentiation of porcine coronavirus from transmissible gastroenteritis virus. Vet. Rec. 1988;122:86–87. doi: 10.1136/vr.122.4.86. [DOI] [PubMed] [Google Scholar]
- Harada K., Kumagai T., Sasahara J. Cytopathogenicity of transmissible gastroenteritis virus in pigs. Nat. Inst. Anim. Hlth. Quart. 1963;3:166–167. [PubMed] [Google Scholar]
- Holm Jensen M. Detection of antibodies against hog cholera virus and bovine viral diarrhoea virus in porcine serum. A comparative examination using CF, PLA and NPLA assays. Acta vet. Scand. 1981;22:85–98. doi: 10.1186/BF03547210. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Honda E., Takahashi H., Okazaki K., Minetoma T., Kumagai T. The multiplication of transmissible gastroenteritis viruses in several cell lines originated from porcine kidney and effects of trypsin on the growth of the viruses. Jap. J. Vet. Sci. 1990;52:217–224. doi: 10.1292/jvms1939.52.217. [DOI] [PubMed] [Google Scholar]
- Jackwood D.J., Kwon H.M., Saif L.J. Molecular differentiation of transmissible gastroenteritis virus and porcine respiratory coronavirus strains. Adv. Exp. Med. Biol. 1995;380:35–41. doi: 10.1007/978-1-4615-1899-0_5. [DOI] [PubMed] [Google Scholar]
- Jones T.O., Paton D.J. Classical transmissible gastroenteritis returns. Vet. Rec. 1996;138:166–167. [PubMed] [Google Scholar]
- Laude H., Van Reeth K., Pensaert M. Porcine respiratory coronavirus: molecular features and virus-host interactions. Vet. Res. 1993;24:125–150. [PubMed] [Google Scholar]
- O'Toole D., Brown I., Bridges A., Cartwright S.F. Pathogenicity of experimental infection with “pneumotropic” porcine coronavirus. Res. Vet. Sci. 1989;47:23–29. doi: 10.1016/S0034-5288(18)31226-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Paton D.J. 2nd edn. Office International des Epizooties; Paris, France: 1992. Transmissible gastroenteritis; pp. 534–542. (Manual of Standards for Diagnostic Tests and Vaccines). [Google Scholar]
- Pensaert M.B., Halterman E.O., Bernstein T. Diagnosis of transmissible gastroenteritis in pigs by means of immunofluorescence. Can. J. Comp. Med. 1968;32:555–561. [PMC free article] [PubMed] [Google Scholar]
- Pensaert M.B., Callebaut P.E., Vergote J. Isolation of a porcine respiratory, non-enteric coronavirus related to transmissible gastroenteritis. Vet. Q. 1986;8:257–260. doi: 10.1080/01652176.1986.9694050. [DOI] [PubMed] [Google Scholar]
- Rowhani A., Maningas M.A., Lile L.S., Daubert S.D., Golino D.A. Development of a detection system for woody plants based on PCR analysis of immobilized virions. Phytopathology. 1995;85:347–352. [Google Scholar]
- Schultze B., Krempl C., Ballesteros M.L., Shaw L., Schauer R., Enjuanes L., Herrler G. Transmissible gastroenteritis coronavirus, but not the related porcine respiratory coronavirus, has a sialic acid (N-glycolylneuramic acid) binding activity. J. Virol. 1996;70:5634–5637. doi: 10.1128/jvi.70.8.5634-5637.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sirinarumitr T., Paul P.S., Kluge J.P., Halbur P.G. In situ hybridization technique for the detection of swine enteric and respiratory coronaviruses, transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV), in formalin-fixed paraffin-embedded tissues. J. Virol. Methods. 1996;56:149–160. doi: 10.1016/0166-0934(95)01901-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stallcup M.R., Washington L.D. Region-specific initiation of mouse mammary tumour virus RNA synthesis by endogenous RNA polymerase II in preparations of cell nuclei. J. Biol. Chem. 1983;258:2802–2807. [PubMed] [Google Scholar]
- Stepanek J., Mesaros E., Pospisil Z. The isolation of the cytopathogenous strains of the originator of transmissible gastroenteritis of pigs in tissue cultures. Vet. Med. Praha. 1969;14:665–674. [Google Scholar]
- Van Nieuwstadt A.P., Boonstra J. Comparison of the antibody response to transmissible gastroenteritis virus and porcine respiratory coronavirus, using monoclonal antibodies to antigenic sites A and X of the S glycoprotein. Am. J. Vet. Res. 1992;53:184–190. [PubMed] [Google Scholar]
- Van Nieuwstadt A.P., Cornelissen J.B., Zetstra T. Comparison of two methods for detection of transmissible gastroenteritis virus in feces of pigs with experimentally induced infection. Am. J. Vet. Res. 1988;49:1836–1843. [PubMed] [Google Scholar]
- Vaughn E.M., Paul P.S. Antigenic and biological diversity among transmissible gastroenetritis virus isolates of swine. Vet. Microbiol. 1993;36:333–347. doi: 10.1016/0378-1135(93)90099-S. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vaughn E.M., Halbur P.G., Paul P.S. Three new isolates of porcine respiratory coronavirus with various pathogenicities and spike (S) gene deletions. J. Clin. Microbiol. 1994;32:1809–1812. doi: 10.1128/jcm.32.7.1809-1812.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vaughn E.M., Halbur P.G., Paul P.S. Use of non-radioactive DNA probes to differentiate porcine respiratory coronavirus and transmissible gastroenteritis virus isolates. J. Vet. Diagn. Invest. 1996;8:241–244. doi: 10.1177/104063879600800216. [DOI] [PubMed] [Google Scholar]
- Wesley R.D., Woods R.D., Hill H.T., Biwer J.D. Evidence for a porcine respiratory coronavirus, antigenically similar to transmissible gastroenteritis virus, in the United States. J. Vet. Diagn. Invest. 1990;2:312–317. doi: 10.1177/104063879000200411. [DOI] [PubMed] [Google Scholar]
- Wesley R.D., Wesley I.V., Woods R.D. Differentiation between transmissible gastroenteritis virus and porcine respiratory using a cDNA probe. J. Vet. Diagn. Invest. 1991;3:29–32. doi: 10.1177/104063879100300106. [DOI] [PubMed] [Google Scholar]
