Comparison of scrapie-associated fibril detection and Western immunoblotting for the diagnosis of natural ovine scrapie

Abstract

Detergent- and proteinase K-treated extracts of grey matter were prepared from four regions of the brains of 106 sheep with scrapie, diagnosed clinically and by the demonstration of spongiform encephalopathy. The extracts were examined by electron microscopy for the presence of scrapie-associated fibrils and by Western immunoblotting for the disease-specific abnormal prion protein (PrPSc). As a diagnostic method, Western immunoblotting proved to be more sensitive than electron microscopy, the detection rates in the 106 sheep being 97 and 91% respectively (medulla), 99 and 76% (cerebellum), 95 and 88% (frontal cerebral cortex) and 93 and 61% (occipital cerebral cortex). Neither fibrils nor PrPSc could be detected in comparable brain extracts from 25 control sheep which had shown no clinical or histopathological evidence of scrapie.