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Scrapie é uma doença que afeta ovinos e caprinos, sendo definida pelo acúmulo de uma isoforma anormal (PrPSc) da proteína priônica celular (PrPC) no Sistema Nervoso Central (SNC) e em tecidos linfoides. Para as técnicas de eleição do diagnóstico de scrapie, imunohistoquímica (IHQ) e western blotting (WB), podem ser utilizados uma vasta gama de anticorpos monoclonais e policlonais comercialmente disponíveis. O objetivo deste estudo foi testar e comparar a eficácia dos anticorpos monoclonais disponíveis comercialmente, F89/160.1.5, F99/97.6.1 e P4, e os anticorpos policlonais R486 e M52, para a identificação da presença da PrPSc em amostras de tecido linfoide e SNC através da técnica de IHQ. Foram utilizadas nas avaliações de IHQ amostras positivas e negativas de cérebro e tonsila palatina de ovinos, cedidas pelo Animal Health Veterinary Laboratory Agency (AHVLA), Reino Unido. Para WB, foram utilizadas amostras de encéfalo, baço, linfonodo, terceira pálpebra e mucosa retal de ovino. As análises IHQ utilizando anticorpos monoclonais e policlonais em amostras positivas de cérebro confirmaram a deposição da PrPSc em neurônios, caracterizada por marcações de aspecto granular intraneural. Na amostra negativa de cérebro, os anticorpos monoclonais não identificaram marcações positivas, o que foi possível verificar ao utilizar os anticorpos policlonais. Testando a amostra positiva de tonsila palatina com os anticorpos monoclonais e policlonais, identificaram-se marcações positivas e, com a amostra negativa, não se observaram marcações com nenhum dos anticorpos, porém observou-se backgroud nos anticorpos monoclonal P4 e policlonais. Apesar dos anticorpos policlonais possuírem valores mais acessíveis, o presente estudo revelou que a utilização dos anticorpos policlonais é inviável, pois a análise pode gerar um resultado falso positivo.Scrapie is a disease that affects sheep and goats and is characterized by the accumulation of an abnormal isoform (PrPSc) of the cellular prion protein, PrPC, in the central nervous system (CNS) and in lymphoid tissues. Detection of PrPSc in these tissues can be attempted by a variety of techniques, including immunohistochemistry (IHC) and western blotting (WB), for which a wide range of monoclonal and polyclonal antibodies are commercially available. The objective of this study was to test and compare the efficacy of monoclonal antibodiesF89/160.1.5, F99/97.6.1, and P4 and polyclonal antibodies M52 and R486 in the detection of PrPSc in lymphoid and CNS tissue samples by using IHC. Positive and negative control samples of sheep brain and tonsils were provided by the Animal Health and Veterinary Laboratories Agency (AHVLA, UK). The IHC examination of CNS samples with both monoclonal and polyclonal antibodies confirmed the granular deposition of PrPSc in the neurons of the positive control tissues. However, while the monoclonal antibodies did not produce positive reactions in the negative controls, the polyclonal antibodies showed some non-specific staining. The testing of positive control tonsil samples with polyclonal and monoclonal antibodies identified positive control-specific reactions, whereas the negative control tissues were IHC-negative with all antibodies, although P4 and the polyclonal antibodies produced some background staining. In summary, although the polyclonal antibodies may be more accessible, their use is not advisable because of possible false positive reactions. The polyclonal antibody M52 was able to identify PrPC in brain and spleen samples by WB but other lymphoid tissues were negative

Monoclonal and polyclonal antibodies for ante- and post-mortem detection of PrPSc in sheep