Evaluation of immunohistochemical detection of prion protein in rectoanal mucosa-associated lymphoid tissue for diagnosis of scrapie in sheep

Objective-To determine the suitability and estimate the sensitivity of an immunohistochemical (IHC) test for disease-associated prion protein (PrP(Sc) in biopsy specimens of rectoanal mucosa-associated lymphoid tissue (RAMALT) for diagnosis of scrapie in sheep. Animals-762 sheep at high risk for having scrapie and indemnified by the National Scrapie Eradication Program. Procedures-The IHC test for PrP(Sc) was applied to 2 RAMALT and 2 third-eyelid biopsy specimens and a postmortem RAMALT specimen from each sheep. Results were compared with those of a reference test in which results for tissues from obex and retropharyngeal lymph nodes, tonsil, or both were considered in parallel. Results-The reference test identified 139 sheep as having scrapie. Biopsy-related complications occurred in 3 sheep. Sensitivity of the IHC test in RAMALT ranged from 85.3% to 89.4%, depending on the anatomic location from which RAMALT was obtained. Results for the test applied to 1 RAMALT specimen were similar to results interpreted in parallel for 2 third-eyelid specimens (sensitivity, 870%). The proportion of inconclusive test results attributable to insufficient lymphoid follicles in biopsy specimens was lower when considering results for 2 RAMALT specimens in parallel (10.1%) than when considering results for 2 third-eyelid specimens in parallel (23.7%). Specimens of RAMALT that were inappropriately collected from an area caudal to the rectoanal interface yielded a high proportion of inconclusive results (33.3% to 50.0%). Conclusions and Clinical Relevance-The IHC test for PrP(Sc) in RAMALT was an effective means of detecting subclinical scrapie in live, high-risk sheep. (Am J Vet Res 2009;70:63-72)