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Journal of Histochemistry and Cytochemistry
Volume 52 (2): 145-151, 2004
Copyright ©The Histochemical Society, Inc.

RAPID COMMUNICATION

Detection of PrPsc on Lymphoid Tissues from Naturally Affected Scrapie Animals : Comparison of Three Visualization Systems

Eva Monleón, Marta Monzón, Paloma Hortells, Antonia Vargas, Cristina Acín and Juan José Badiola

National Reference Centre for TSEs, University of Zaragoza, Zaragoza, Spain

Correspondence to: Juan José Badiola Díez, National Reference Centre for Transmissible Spongiform Encephalopathies, Dept. of Animal Pathology of the Veterinary Faculty, C/ Miguel Servet 177, 50013 Zaragoza. Spain. E-mail: badiola{at}unizar.es

We assessed three different visualization systems used routinely in research and diagnosis of transmissable spongiform encephalopathies (TSEs) to demonstrate whether the methodology applied to immunohistochemical (IHC) examination may alter the results concerning detection of prion protein (PrPsc) in the lymphoreticular system (LRS): avidin–biotin–peroxidase (Vectastain ABC kit; Vector), Envision (DAKO), and catalyzed signal amplification (CSA; DAKO). The study aimed to determine which of these showed the highest sensitivity, with the hope of providing an accurate tool for pathogenesis and preclinical diagnosis research in TSEs. Histological sections from palatine tonsils, spleen, GALT (ileum and ileocecal valve), and lymph nodes from sheep belonging to a Spanish scrapie-positive flock were processed by IHC using L42 as primary antibody. As substrate chromogen, diaminobenzidine was used, and all slides were subjectively assessed by light microscopy. A further study using an image analyzer software system was carried out to confirm that the conclusion provided by microscopic examination was objective. The CSA system showed the highest sensitivity in all cases, increasing both variables assessed: the number of follicles that were PrPsc-positive was detected as well as the intensity of immunostaining in each of them. (J Histochem Cytochem 52:145–151, 2004)

Key Words: TSEs • scrapie • immunohistochemistry • lymphoid tissue • visualization systems


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