Immunohistochemical Detection of Disease-Associated Prion Protein in the Intestine of Cattle Naturally Affected with Bovine Spongiform Encephalopathy by Using an Alkaline-Based Chemical Antigen Retrieval Method

Abstract

An alkaline-based chemical antigen retrieval pretreatment step was used to enhance immunolabeling of disease-associated prion protein (PrP^Sc) in formalin-fixed and paraffin-embedded tissue sections from cattle naturally affected with bovine spongiform encephalopathy (BSE). The modified chemical method used in this study amplified the PrP^Sc signal by unmasking PrP^Sc compared with the normal cellular prion protein. In addition, this method reduced nonspecific background immunolabeling that resulted from the destruction of the residual normal cellular form of prion protein, and reduced the treatment time compared with the usual autoclave pretreatment step. Immunolabeled PrP^Sc was thereby clearly detected in the myenteric plexus of the ileum in naturally occurring BSE cattle.