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Journal of Clinical Microbiology, September 2000, p. 3254-3259, Vol. 38, No. 9
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.
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
*
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.
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