Skip to main content
Log in

Comparison of Immunoperoxidase Plaque Staining and Neutralizing Tests for Canine Distemper Virus

  • Published:
Veterinary Research Communications Aims and scope Submit manuscript

Abstract

The use of immunoperoxidase plaque staining (IP) in the antibody test for canine distemper virus (CDV) and its diagnostic value were investigated. Antibody titres by the IP test and neutralizing test (NT) against CDV were compared in 114 specific pathogen-free (SPF) dogs, including 84 CDV-vaccinated and 30 unimmunized dogs. All of the unimmunized dogs, which had negative NT titres, also showed negative IP titres, and all of the CDV-vaccinated dogs, which had positive NT titres, showed positive IP titres. Furthermore, the IP titres showed similar changes with time after vaccination to the NT titres in five CDV-vaccinated SPF dogs. These findings indicate that the IP test has similar specificity to the NT.

Next, the IP and NT titres were compared in 190 clinically suspected cases of canine distemper (CD), including 107 dogs with neurological signs and 83 dogs without neurological signs, and also in 50 clinically healthy dogs. Among the suspected cases of CD, of 46 dogs with neurological signs and 29 dogs without neurological signs, and which had negative NT titres, 33 (72%) and 19 (66%), respectively, showed positive IP titres. Many suspected cases of CD showed a high IP titre despite a low NT titre. In contrast, among healthy dogs, none of 9 NT-negative dogs showed positive IP titres.

The difference in the IP and NT titres (log10 [reciprocal of IP titre] – log10 [reciprocal of NT titre]) was measured to investigate the agreement between IP test and NT. The differences were 1.488±0.867, 1.245±0.833, 0.595±0.351 and 0.594±0.372, respectively, in the suspected cases of CD with neurological signs, the suspected cases of CD without neurological signs, the healthy dogs and the SPF dogs. The differences in the suspected cases of CD were significantly high compared to those for the SPF dogs (p<0.001).

Furthermore, in 8 dogs diagnosed as cases of CD from their paired sera, whose initial sera showed a markedly low NT titre compared to the IP titre, 5 dogs without neurological signs developed neurological signs later.

These findings suggest that the IP test can detect CD during its early phases and in dogs with low antibody levels, undetectable by NT, and so would be useful for serological diagnosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

REFERENCES

  • Appel, M.J.G., 1969. Pathogenesis of canine distemper. American Journal of Veterinary Research, 30, 1167–1182

    Google Scholar 

  • Appel, M. and Robson, D.S., 1973. A microneutralization test for canine distemper virus. American Journal of Veterinary Research, 34, 1459–1463

    Google Scholar 

  • Bernard, S.L., Shen, D.T. and Gorham, J.R., 1982. Antigen requirements and specificity of enzyme-linked immunosorbent assay for detection of canine IgG against canine distemper viral antigens. American Journal of Veterinary Research, 43, 2266–2269

    Google Scholar 

  • Black, J.W., 1983. Single serum-sample diagnosis of canine viral diseases. Veterinary Medicine, 78, 1393–1396

    Google Scholar 

  • Bland, J.M. and Altman, D.G., 1986. Statistical methods for assessing agreement between two methods of clinical measurement. Lancet, 1, 307–310

    Google Scholar 

  • Blixenkrone-Moller, M., Pedersen, I.R., Appel, M.J. and Griot, C., 1991. Detection of IgM antibodies against canine distemper virus in dog and mink sera employing enzyme-linked immunosorbent assay (ELISA). Journal of Veterinary Diagnostic Investigation, 3, 3–9

    Google Scholar 

  • Greene, C.E., 1984. Canine distemper. In: C.E. Greene (ed.), Clinical Microbiology and Infectious Diseases of the Dog and Cat, (W.B. Saunders, Philadelphia), 386–405

    Google Scholar 

  • Handy, F.M., Colgrove, G.S., Rodrigues, E.M., Snyder, M.L. and Stewart, W.C., 1981. Field evaluation of enzyme-linked immunosorbent assay for detection of antibody to African swine fever virus. American Journal of Veterinary Research, 42, 1441–1443

    Google Scholar 

  • Kai, K., Yukimune, M., Murata, T., Uzuka, Y., Kanoe, M. and Matsumoto, H., 1992. Humoral immune responses of cats to feline infectious peritonitis virus infection. Journal of Veterinary Medical Science, 54, 501–507

    Google Scholar 

  • Krakowka, S., Olsen, R., Confer, A., Koestner, A. and McCullough, B., 1975. Serologic response to canine distemper viral antigens in gnotobiotic dogs infected with canine distemper virus. Journal of Infectious Diseases, 132, 384–392

    Google Scholar 

  • Kyhse-Andersen, J., 1984. Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. Journal of Biochemical and Biophysical Methods, 10, 203–209

    Google Scholar 

  • Laemmli, U.K., 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London), 227, 680–685

    Google Scholar 

  • Liu, C. and Coffin, D.L., 1957. Studies on canine distemper infection by means of fluorescein-labeled antibody 1. The pathogenesis, pathology, and diagnosis of the disease in experimentally infected ferrets. Virology, 3, 115–131

    Google Scholar 

  • Miele, J.A. and Krakowka, S., 1983. Antibody responses to virion polypeptides in gnotobiotic dogs infected with canine distemper virus. Infection and Immunity, 41, 869–871

    Google Scholar 

  • Noon, K.F., Rogul, M., Binn, L.N., Keefe, T.J., Marchwicki, R.H. and Appel, M.J., 1980. Enzyme-linked immunosorbent assay for evaluation of antibody to canine distemper virus. American Journal of Veterinary Research, 41, 605–609

    Google Scholar 

  • Pan, I.C., Huang, T.S. and Hess, W.R., 1982. New method of antibody detection by indirect immunoperoxidase plaque staining for serodiagnosis of African swine fever. Journal of Clinical Microbiology, 16, 650–655

    Google Scholar 

  • Reed, L.T. and Muench, H., 1938. A simple method of estimating fifty percent end points. American Journal of Hygiene, 27, 493–497

    Google Scholar 

  • Rima, B.K., Baczko, K., Imagawa, D.T. and Meulen, V.T., 1987. Humoral immune response in dogs with old dog encephalitis and chronic distemper meningoencephalitis. Journal of General Virology, 68, 1723–1735

    Google Scholar 

  • Rima, B.K., Duffy, N., Mitchell, W.J., Summers, B.A. and Appel, M.J.G., 1991. Correlation between humoral immune responses and presence of virus in the CNS in dogs experimentally infected with canine distemper virus. Archives of Virology, 121, 1–8

    Google Scholar 

  • Sato, K., Tanaka, Y., Kurogi, H., Tokuhisa, S., Numba, K., Inaba, Y. and Matumoto, M., 1988. Detection of antibody to pseudorabies virus in swine sera by indirect immunoperoxidase plaque staining. Journal of Clinical Microbiology, 26, 79–81

    Google Scholar 

  • Shetty, N.P., Raj, I.S. and Macaden, R.S., 1990. Non-specific reactions in enzyme linked immunosorbent assays for serum antibody to Entamoeba histolytica and Giardia lamblia in non-endemic areas. Journal of Clinical Pathology, 43, 950–952

    Google Scholar 

  • Tajima, T., Yoshizaki, S., Nakata, E., Tohya, Y., Ishiguro, S., Fujikawa, Y. and Sugii, S., 1998. Production of a monoclonal antibody reacted broadly with feline calicivirus field isolates. Journal of Veterinary Medical Science, 60, 155–160

    Google Scholar 

  • 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

    Google Scholar 

  • Warren, R.C., White, L.O., Mohan, S. and Richardson, M.D., 1979. The occurrence and treatment of false positive reactions in enzyme-linked immunosorbent assays (ELISA) for the presence of fungal antigens in clinical samples. Journal of Immuno logical Methods, 28, 177–186

    Google Scholar 

  • Yamaguchi, K., Matutes, E., Kiyokawa, T., Nishimura, Y., Ishii, T., Takatsuki, K. and Catovsky, D., 1988. Comparison of immunoperoxidase staining with indirect immunofluorescence, ELISA, and western blotting assays for detecting anti-HTLV-1 antibodies in systemic lupus erythematosus. Journal of Clinical Pathology, 41, 57–61

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Soma, T., Ishii, H., Hara, M. et al. Comparison of Immunoperoxidase Plaque Staining and Neutralizing Tests for Canine Distemper Virus. Vet Res Commun 25, 311–325 (2001). https://doi.org/10.1023/A:1010682726245

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1010682726245

Navigation