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Clinical and Diagnostic Laboratory Immunology, September 2005, p. 1050-1056, Vol. 12, No. 9
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.9.1050-1056.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Enzyme-Linked Immunosorbent Assays for Detection of Equine Antibodies Specific to Sarcocystis neurona Surface Antigens{dagger}

Jessica S. Hoane,1 Jennifer K. Morrow,2 William J. Saville,3 J. P. Dubey,4 David E. Granstrom,4 and Daniel K. Howe1*

Department of Veterinary Science, University of Kentucky, 108 Gluck Equine Research Center, Lexington, Kentucky 40546-0099,1 Equine Biodiagnostics/IDEXX, 1501 Bull Lea Road, Suite 104, Lexington, Kentucky 40511,2 Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, 1900 Coffey Road, Columbus, Ohio 43210-1092,3 United States Department of Agriculture, Agricultural Research Service, Animal and Natural Resources Institute, Beltsville, Maryland 20705-23504

Received 17 March 2005/ Returned for modification 25 May 2005/ Accepted 10 June 2005

Sarcocystis neurona is the primary causative agent of equine protozoal myeloencephalitis (EPM), a common neurologic disease of horses in the Americas. We have developed a set of enzyme-linked immunosorbent assays (ELISAs) based on the four major surface antigens of S. neurona (SnSAGs) to analyze the equine antibody response to S. neurona. The SnSAG ELISAs were optimized and standardized with a sample set of 36 equine sera that had been characterized by Western blotting against total S. neurona parasite antigen, the current gold standard for S. neurona serology. The recombinant SnSAG2 (rSnSAG2) ELISA showed the highest sensitivity and specificity at 95.5% and 92.9%, respectively. In contrast, only 68.2% sensitivity and 71.4% specificity were achieved with the rSnSAG1 ELISA, indicating that this antigen may not be a reliable serological marker for analyzing antibodies against S. neurona in horses. Importantly, the ELISA antigens did not show cross-reactivity with antisera to Sarcocystis fayeri or Neospora hughesi, two other equine parasites. The accuracy and reliability exhibited by the SnSAG ELISAs suggest that these assays will be valuable tools for examining the equine immune response against S. neurona infection, which may help in understanding the pathobiology of this accidental parasite-host interaction. Moreover, with modification and further investigation, the SnSAG ELISAs have potential for use as immunodiagnostic tests to aid in the identification of horses affected by EPM.

* Corresponding author. Mailing address: 108 Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, KY 40546-0099. Phone: (859) 257-4757, ext. 81113. Fax: (859) 257-8542. E-mail: dkhowe2{at}uky.edu.

{dagger} Published as Kentucky Agricultural Experiment Station article no. 05-14-023.

Clinical and Diagnostic Laboratory Immunology, September 2005, p. 1050-1056, Vol. 12, No. 9
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.9.1050-1056.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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