Abstract
One complement-fixing (C-MAb) and three complement-dependent neutralizing monoclonal antibodies (N-MAbs) were raised against Hisar-90-7 equine herpesvirus-1 (EHV-1) strain. The target antigen of the C-MAb (2A5) and two of the N-MAbs (1H6, 9C4) was identified as a 140 kDa polypeptide in Western blotting. The target antigen of N-MAb (9C6) could not be identified. Purified polypeptides of five EHV-1 strains isolated from different regions and at different times gave intense bands at 140 kDa when reacted with N-MAb (1H6) in Western blots. In sandwich ELISA, all four MAbs captured the viral antigen from clinical materials, giving a reliable and rapid diagnosis of EHV-1 infection in equines.
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REFERENCES
Allen, G.P. and Yeargan, M.R., 1987. Use of lambda gt-II monoclonal antibodies to map the genes for the six major glycoproteins of equine herpes virus-1. Journal of Virology, 61, 2454-2461
Allen, G.P., Coogle, L.D., Ostlund, E.N. and Yeargan, M.R., 1991. Molecular dissection of two major equine herpes virus-1 glycoprotein antigens (gB and gC) that elicit humoral immune responses in the horse. In: W. Plowright, P.D. Rossddale and J.F. Wade (eds), Equine Infectious Diseases, VI, (Cambridge University Press, Cambridge), 181-193
Allen, G.P., Yeargan, M.R. and Coogle, L.D., 1988. Equid herpes virus-1 glycoprotein 13 (gp13): epitope analysis, gene structure and expression in E. coli. In: D.G. Powell (ed.), Equine Infectious Diseases V: Proceedings of the Fifth International Conference on Equine Infectious Diseases, (University Press of Kentucky, Lexington), 103-110
Barlow, D.J., Edwards, M.S. and Thornton, J.M., 1986. Continuous and discontinuous protein antigenic determinants. Nature, 322, 747-748
Campbell, A.M., 1987. Fusion procedure: selection and cloning. In: R.H. Budron (ed.), Monoclonal Antibody Technology, (Elsevier Science, Amsterdam), 120-164
Collins, J.K., Butcher, A.C., Riegel, C.A., McGrane, V., Blair, C.D., Teramoto, Y.A. and Winston, S., 1984. Neutralizing determinants defined by monoclonal antibodies on polypeptides specified by bovine herpes virus-1. Journal of Virology, 52, 403-409
Cottral, E.G., 1978. Manual of Standardized Methods for Microbiology, (Cornell University Press, Ithaca, NY), 75-85
Crabb, B.S. and Studdert, M.J., 1993. Epitopes of glycoprotein G of equine herpesviruses 4 and 1 located near the C termini elicit type-specific antibody responses in the natural host. Journal of Virology, 6332-6338
Crabb, B.S., Allen, G.P. and Studdert, M.J., 1991. Characterization of the major glycoproteins of equine herpes virus 4 and 1 and asinine herpes virus 3 using monoclonal antibodies. Journal of General Virology, 72, 2075-2082
Engvall, E. and Perlmann, P., 1972. Enzyme-linked immunosorbent assay. ELISA III. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes. Journal of Immunology, 109, 129-135
Kohler, G. and Milstein, C., 1975. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature, 256, 495-497
Okazaki, K., Kumagai, T., Honda, E. and Okazaki, K., 1990. The immunodominant glycoprotein complex of equid herpesvirus-1 (EHV-1) and the counterpart of EHV-4. Japanese Journal of Veterinary Science, 52, 1127-1130. (Seen in Veterinary Bulletin (1991) 61, Abstr. no. 3369)
Russell, P.K., Brandt, W.E. and Dalrymple, J.M., 1980. Chemical and antigenic structure of flaviviruses. In: R.W. Schlesinger (ed.), The Togaviruses-Biology, Structure, Replication, (Academic Press, New York), 503-529
Schlesinger, J.J., Brandriss, M.W. and Monath, T.P., 1983. Monoclonal antibodies distinguish between wild and vaccine strains of yellow fever virus by neutralization, hemagglutination inhibition and immune precipitation of the virus envelope protein. Virology, 125, 8-17
Shimizu, M., Satou, K. and Nishioka, N., 1989. Monoclonal antibodies with neutralizing activity to equine herpesvirus-1. Archives of Virology, 104, 169-174
Sinclair, R. and Mumford, J.A., 1992. Rapid detection of equine herpesvirus type-1 antigens in nasal swab specimens using an antigen capture enzyme-linked immunosorbent assay. Journal of Virological Methods, 39, 299-310
Sinclair, R., Cook, R.F. and Mumford, J.A., 1989. The characterization of neutralizing and non-neutralizing monoclonal antibodies against equid herpesvirus type 1. Journal of General Virology, 70, 455-459
Tewari, S.C., Verma, P.C. and Bhargava, D.N., 1994. Experimental equine herpes virus-1 infection of foetus in a pregnant donkey mare. Indian Veterinary Journal, 71, 213-214
Thomson, G.R., Mumford, J.A., Campbell, J., Griffith, L. and Clapham, P., 1976. Serological detection of equid herpes virus-1 infections in the respiratory tract. Equine Veterinary Journal, 8, 58-65
Tizard, I.R., 1995. Immunology: An Introduction, 4th edn, (Saunders College Publishing, New York)
Towbin, H., Staehelin, T. and Gordon, J., 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences of the USA, 76, 4350-4354
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Singh, B., Yadav, M. & Tewari, S. Neutralizing and Complement-fixing Monoclonal Antibodies as an Aid to the Diagnosis of Equine Herpesvirus-1 Infection. Vet Res Commun 25, 675–686 (2001). https://doi.org/10.1023/A:1012747331088
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DOI: https://doi.org/10.1023/A:1012747331088