Journal article
Serological diagnosis of bovine neosporosis by Neospora caninum monoclonal antibody-based competitive inhibition enzyme-linked immunosorbent assay
Journal of clinical microbiology, Vol.34(6), pp.1423-1428
06/1996
Handle:
https://hdl.handle.net/2376/116054
PMCID: PMC229036
PMID: 8735092
Abstract
Neospora caninum, a protozoan parasite closely related to Toxoplasma gondii, causes abortion and congenital infection in cattle. To investigate specific methods of antemortem diagnosis, the antibody responses of infected cows were evaluated by immunoblot assay and competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA) by using a monoclonal antibody (MAb), MAb 4A4-2, against N. caninum tachyzoites. MAb 4A4-2 bound diffusely to the exterior surface of N. caninum tachyzoites and recognized a single 65-kDa band in immunoblots. MAb 4A4-2 was unreactive to antigens of two closely related apicomplexan protozoa, Toxoplasma gondii and Sarcocystis cruzi. Binding of MAb 4A4-2 was inhibited by mild periodate treatment of N. caninum antigen, demonstrating the carbohydrate nature of the epitope. Immunoblot analysis of N. caninum tachyzoite antigens with sera from cows with confirmed Neospora-induced abortion revealed at minimum 14 major antigens ranging from 11 to 175 kDa. Although the recognized antigens varied from cow to cow, antigens of 116, 65, and 25 kDa were detected in all cows with abortion confirmed to be caused by N. caninum. The binding of MAb 4A4-2 to N. caninum tachyzoite antigen was consistently inhibited by sera from Neospora-infected cows in a CI-ELISA format and was not inhibited by sera from Neospora antibody-negative cows. Furthermore, sera from cattle experimentally infected with T. gondii, S. cruzi, Sarcocystis hominis, or Sarcocystis hirsuta, which had cross-reactive antibodies recognizing multiple N. caninum antigens by immunoblot assay, did not inhibit binding of MAb 4A4-2 in the CI-ELISA. Thus, MAb 4A4-2 binds a carbohydrate epitope on a single N. caninum tachyzoite surface antigen that is recognized consistently and specifically by Neospora-infected cattle.
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Details
- Title
- Serological diagnosis of bovine neosporosis by Neospora caninum monoclonal antibody-based competitive inhibition enzyme-linked immunosorbent assay
- Creators
- T V Baszler - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164-7040, USA. baszlert@vetmed.wsu.eduD P Knowles - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164-7040, USA. baszlert@vetmed.wsu.eduJ P Dubey - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164-7040, USA. baszlert@vetmed.wsu.eduJ M Gay - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164-7040, USA. baszlert@vetmed.wsu.eduB A Mathison - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164-7040, USA. baszlert@vetmed.wsu.eduT F McElwain - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164-7040, USA. baszlert@vetmed.wsu.edu
- Publication Details
- Journal of clinical microbiology, Vol.34(6), pp.1423-1428
- Academic Unit
- Veterinary Clinical Sciences, Department of; Veterinary Microbiology and Pathology, Department of; Paul G. Allen School for Global Animal Health
- Identifiers
- 99900547782801842
- Language
- English
- Resource Type
- Journal article