Monoclonal antibodies (mAb) to the transmissible gastroenteritis virus (TGEV) nucleoprotein (N) and membrane protein (M) were prepared and used for the comparative assessment of three blocking ELISA variants to detect TGEV. The competitive blocking ELISA format showed the highest sensitivity, allowing detection of 10(3) TCID50 TGEV/ml in culture medium. Ninety-nine porcine field faecal samples obtained from 37 herds affected with diarrhoea were examined, and various TGEV levels were found in nine samples from six herds. However, only in three samples were significant TGEV concentrations demonstrated. The relationship between incidence of TGEV gastroenteritis and the spread of porcine respiratory coronavirus infection in pig farms is discussed.

Veterinary Research Institute Hudcova 70 621 32 Brno Czech Republic

Zobrazit více v PubMed

Aynaud, J. M. , and Bottreau E., 1984: Transmissible gastroenteritis of swine: stability of coronavirus in gastric and intestinal contents. Ann. Rech. Vet. 15, 359–364. PubMed

Ballesteros, M. L. , Sanchez C. M., and Enjuanes L., 1997: Two amino acid changes at the N‐terminus of transmissible gastroenteritis coronavirus spike protein result in the loss of enteric tropism. Virology 227, 378–388. PubMed PMC

Bernard, S. , Bottreau E., Aynaud J. M., Have P., and Szymansky J., 1989: Natural infection with the porcine respiratory coronavirus induces protective lactogenic immunity against transmissible gastroenteritis. Vet. Microbiol. 21, 1–8. PubMed PMC

Boorsma, D. M. , and Streefkerk J. G., 1979: Periodate or glutaraldehyde for preparing peroxidase conjugates? J. Immunol. Methods 30, 245–255. PubMed

Britton, P. , Mawditt K. L., and Page K. W., 1991: The cloning and sequencing of the virion protein genes from a British isolate of porcine respiratory coronavirus: comparison with transmissible gastroenteritis virus genes. Virus Res. 21, 181–198. PubMed PMC

Carvajal, A. , Lanza I., Diego R., Rubio P., and Cármenes P., 1995: Evaluation of a blocking ELISA using monoclonal antibodies for the detection of porcine epidemic diarrhea virus and its antibodies. J. Vet. Diagn. Invest. 7, 60–64. PubMed

Chae, C. , Kim O., Choi C., Min K., Cho W.‐S., Kim J., and Tai J. H., 2000: Prevalence of porcine epidemic diarrhoea virus and transmissible gastroenteritis virus infection in Korean pigs. Vet. Rec. 18, 606–608. PubMed

Constantini, V. , Lewis P., Alsop J., Templeton C., and Saif L. J., 2004: Respiratory and fecal shedding of porcine respiratory coronavirus (PRCV) in sentinel weaned pigs and sequence of the partial S‐gene of the PRCV isolates. Arch. Virol. 149, 957–974. PubMed PMC

Cox, E. , Hooyberghs J., and Pensaert M. B., 1990a: Sites of replication of a porcine respiratory coronavirus related to transmissible gastroenteritis virus. Res. Vet. Sci. 48, 165–169. PubMed PMC

Cox, E. , Pensaert M. B., Callebaut P., and van Deun K., 1990b: Intestinal replication of a porcine respiratory coronavirus closely related antigenically to the enteric transmissible gastroenteritis virus. Vet. Microbiol. 23, 237–243. PubMed PMC

Cox, E. , Pensaert M. B., Hooyberghs J., and van Deun K., 1990c: Sites of replication of a porcine respiratory coronavirus in 5‐week‐old pigs with or without maternal antibodies. Adv. Exp. Med. Biol. 276, 429–433. PubMed

Galfrè, G. , and Milstein C., 1981: Preparation of monoclonal antibodies: strategies and procedures. Methods Enzymol. 73, 3–46. PubMed

Hofmann, M. , and Wyler R., 1990: Enzyme‐linked immunosorbent assay for the detection of porcine epidemic diarrhea coronavirus antibodies in swine sera. Vet. Microbiol. 21, 263–273. PubMed PMC

Kim, B. H. , and Cho K. H., 1998: Transmissible gastroenteritis of pigs in southeastern Korea in 1994–1996: prevalence and factors associated with the infection In: Done S., Thomson S., and Varley M. (eds) Proceedings of the 15th IPVS Congress, Birmingham, UK, 5–9 July, p. 146. Nottingham University Press, Manor farm, Nottingham.

Kim, S. Y. , Song D. S., and Park B. K., 2001: Differential detection of transmissible gastroenteritis virus and porcine epidemic diarrhea virus by duplex RT‐PCR. J. Vet. Diagn. Invest. 13, 516–520. PubMed

Laemmli, U. K. , 1970: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685. PubMed

Lanza, I. , Shoup D. I., and Saif L. J., 1995: Lactogenic immunity and milk antibody isotypes to transmissible gastroenteritis virus in sows exposed to porcine respiratory coronavirus during pregnancy. Am. J. Vet. Res. 56, 739–748. PubMed

Li, C. Y. , Ziesmer S. C., and Lazcano‐Villareal O., 1987: Use of azide and hydrogen peroxide as an inhibitor for endogeneous peroxidase in the immunoperoxidase method. J. Histochem. Cytochem. 35, 1457–1460. PubMed

Moeremans, M. , Daniels G., and de Mey J., 1985: Sensitive colloidal (gold or silver) staining of protein blots on nitrocellulose membranes. Anal. Biochem. 145, 315–321. PubMed

van Nieuwstadt, A. P. , Cornelissen J. B., and Zetstra T., 1988: Comparison of two methods for detection of transmissible gastroenteritis virus in feces of pigs with experimentally induced infection. Am. J. Vet. Res. 49, 1836–1843. PubMed

Paton, D. , Ibata G., Sands J., and McGoldrick A., 1997: Detection of transmissible gastroenteritis virus by RT‐PCR and differentiation from porcine respiratory coronavirus. J. Virol. Methods 66, 303–309. PubMed PMC

Pritchard, G. C. , Paton D. J., Wibberley G., and Ibata G., 1999: Transmissible gastroenteritis and porcine epidemic diarrhoea in Britain. Vet. Rec. 144, 616–618. PubMed

Rasschaert, D. , Duarte M., and Laude H., 1990: Porcine respiratory coronavirus differs from transmissible gastroenteritis virus by a few genomic deletions. J. Gen. Virol. 71, 2599–2607. PubMed

Rodák, L. , Šmíd B., Nevoránková Z., Smítalová R., and Valíček L., 2004: Verification of sensitivity and specificity of group A rotavirus detection in piglets faeces with monoclonal blocking ELISA methods. J. Vet. Med. B 51, 160–165. PubMed PMC

Rodák, L. , Valíček L. Šmíd B., and Nevoránková Z., 2005: An ELISA optimized for porcine epidemic diarrhoea virus detection in faeces. Vet. Microbiol. 105, 9–17. PubMed PMC

Saif, L. J. , 1993. Coronavirus immunogens. Vet. Microbiol. 37, 285–297. PubMed PMC

Saif, L. J , van Cott J. L., and Brim T. A., 1994: Immunity to transmissible gastroenteritis virus and porcine respiratory coronavirus infections in swine. Vet. Immunol. Immunopathol. 43, 89–97. PubMed PMC

Šesták, K. , Lanza I., Park S. K., Weilnau P. A., and Saif L. J., 1996: Contribution of passive immunity to porcine respiratory coronavirus to protection against transmissible gastroenteritis virus challenge exposure in suckling pigs. Am. J. Vet. Res. 57, 664–671. PubMed

Simkins, R. A. , Weilnau P. A., van Cott J., Brim T. A., and Saif L. J., 1993: Competition ELISA, using monoclonal antibodies to the transmissible gastroenteritis virus (TGEV) S protein, for serologic differentiation of pigs infected with TGEV or porcine respiratory coronavirus. Am. J. Vet. Res. 54, 254–259. PubMed

Šmíd, B. , Valíček L., Rodák L., and Kudrna J., 1993: Electron microscopic demonstration of porcine epidemic diarrhea virus in the Czech Republic. Vet. Med. Czech. 38, 333–341 (in Czech). PubMed

Štěpánek, J. , Menšík J., Rozkošný V., and Mesároš E., 1972: Transmissible gastroenteritis of swine (TGE). Physicochemical and biological properties of strains isolated in Czechoslovakia. Acta Vet. (Brno), 41, 435–445.

Utiger, A. , Tobler K., Bridgen A., Suter M., Singh M., and Ackermann M., 1995: Identification of proteins specified by porcine epidemic diarrhoea virus In: Talbot P. J., and Levy G. A. (eds), Corona – and Related Viruses, pp. 287–290. Plenum Press, New York. PubMed