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Journal of Histochemistry and Cytochemistry, Vol. 51, 15-18, January 2003, Copyright © 2003, The Histochemical Society, Inc.

RAPID COMMUNICATION

Detection of PrPsc in Samples Presenting a Very Advanced Degree of Autolysis (BSE Liquid State) by Immunocytochemistry

Eva Monleóna, Marta Monzóna, Paloma Hortellsa, Antonia Vargasa, and Juan J. Badiolaa
a National Reference Centre for Transmissible Spongiform Encephalopathies, University of Zaragoza, Zorgoza, Spain

Correspondence to: Juan J. Badiola, National Reference Centre for Transmissible Spongiform Encephalopathies, Dept. of Pathological Anatomy of the School of Veterinary Medicine, Miguel Servet 177, 50013 Zaragoza, Spain. E-mail: badiola@posta.unizar.es

Although detection of the abnormal isoform of prion protein (PrPsc), the specific feature of transmissable spongiform encephalopathies (TSEs), has been previously demonstrated on formalin-fixed autolytic tissue, no samples with autolysis as severe as tested here (i.e., liquid state) have previously been tested. It is inevitable that a small but significant proportion of brains, especially in summer due to delays in postmortem examination, undergo an extremely severe autolysis that makes samples unsuitable for diagnosis by conventional techniques. In this study, 25 bovine samples were diagnosed by applying immunocytochemistry on the corresponding liquid fraction. Four additional portions of brainstem (positive and negative sheep and cattle) were subjected to one of the autolysis regimens at 56C or environmental conditions for up to 80 days and were analyzed with the same methodology. No abnormal protein could be detected in any of the control animals. PrPsc accumulation was observed by immunocytochemistry in all cases that were positive by either immunohistochemistry on the corresponding filtrates or by Prionics Western blotting, showing an excellent agreement between the methodology assessed and these routine techniques. The results of this study demonstrate immunocytochemistry as a useful tool for diagnosis in liquid-state samples, solving a most relevant problem in BSE and scrapie epidemiology. (J Histochem Cytochem 51:15–18, 2003)

Key Words: autolysis, liquid state, BSE, scrapie, immunocytochemistry, PrPsc


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