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Journal of Clinical Microbiology, September 2000, p. 3254-3259, Vol. 38, No. 9
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Preclinical Diagnosis of Scrapie by Immunohistochemistry of Third Eyelid Lymphoid Tissue

K. I. O'Rourke,1,* T. V. Baszler,2,3 T. E. Besser,2 J. M. Miller,4 R. C. Cutlip,4 G. A. H. Wells,5 S. J. Ryder,5 S. M. Parish,6 A. N. Hamir,4 N. E. Cockett,7 A. Jenny,8 and D. P. Knowles1,2

Animal Disease Research Unit, Animal Research Service, U.S. Department of Agriculture,1 Department of Veterinary Microbiology and Pathology2 and Department of Clinical Veterinary Sciences,6 College of Veterinary Medicine, Washington State University, and Washington Animal Disease Diagnostic Laboratory,3 Pullman, Washington; National Animal Disease Center, Agricultural Research Service,4 and National Veterinary Services Laboratories, Animal and Plant Health Inspection Service,8 U.S. Department of Agriculture, Ames, Iowa; Veterinary Laboratories Agency, Ministry of Agriculture, Fisheries, and Food, Surrey, United Kingdom5; and Department of Animal, Dairy, and Veterinary Sciences, Utah State University, Logan, Utah7

Received 13 January 2000/Returned for modification 18 April 2000/Accepted 1 June 2000

Ovine scrapie is a member of the transmissible spongiform encephalopathies (TSEs), a heterogeneous family of fatal neurologic disorders characterized by deposition of an abnormal isoform (prion protein [PrP] PrP-Sc) of a cellular sialoglycoprotein in neural tissue. PrP-Sc is detectable in some lymphoid tissues of infected sheep months or years before development of clinical disease. Detection of PrP-Sc in these tissues is the basis for live-animal testing. In this study, we characterize the performance of a preclinical diagnostic test for ovine scrapie based on a monoclonal antibody (MAb)-based immunohistochemistry assay of nictitating membrane ("third eyelid")-associated lymphoid tissue. The results of third eyelid immunohistochemistry assay agreed with the scrapie status of the sheep for 41 of 42 clinical suspects with confirmed scrapie and 174 of 175 sheep without scrapie. Third eyelid sampling agreed with the scrapie status for 36 of 41 clinically normal sheep positive for PrP-Sc immunostaining of brain tissue, including 27 sheep with positive biopsy specimens that progressed to clinical disease with confirmed scrapie 3 to 20 months after biopsy. The assay used MAb F89/160.1.5, which binds to residues 142 to 145 of ovine PrP. This antibody can be used in combination with MAb F99/97.6.1, which binds to residues 220 to 225. One or both MAbs in this cocktail recognize PrP sequences conserved in most mammalian species in which natural TSEs have been reported. Immunohistochemistry assay of routinely formalin-fixed lymphoid tissues with a cocktail of pan-specific MAbs is a practical, readily standardized live-animal and preclinical test for ovine scrapie.

* Corresponding author. Mailing address: USDA, ARS, ADRU, 3003 ADBF, P.O. Box 646630, Pullman, WA 99164-6630. Phone: (509) 335-6020. Fax: (509) 335-8328. E-mail: korourke{at}vetmed.wsu.edu.

Journal of Clinical Microbiology, September 2000, p. 3254-3259, Vol. 38, No. 9
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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