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Year 2008 - Volume 28, Number 12
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Padronização da técnica de imuno-histoquímica para raiva em amostras de tecido do sistema nervoso central de bovinos fixadas em formol e emblocadas em parafina, p.627-632
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ABSTRACT.- Pedroso P.M.O., Pescador C.A., Bandarra P.M., Raymundo D.L., Borba M.R., Wouters F., Bezerra Jr P.S. & Driemeier D. 2008. [Standardization of immunohistochemistry technique for detection of rabies virus in formalin-fixed and paraffin-embedded tissue samples from central nervous system of cattle.] Padronização da técnica de imuno-histoquímica para raiva em amostras de tecido do sistema nervoso central de bovinos fixadas em formol e emblocadas em parafina. Pesquisa Veterinária Brasileira 28(12):627-632. Setor de Patologia Veterinária, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Av. Bento Gonçalves 9090, Porto Alegre, RS 91540-000, Brazil. E-mail: davetpat@ufrgs.br
For standardization of the rabies immunohistochemistry technique, five samples of central nervous system (CNS) of cattle naturally infected with rabies virus were examined. One polyclonal antibody and two monoclonal antibodies were used. The following reagents were evaluated for antigen retrieval: XIV protease, proteinase K and citrate buffer (pH 6.0) boiling at 100°C during 15 minutes in bain-marie. Detection of rabic antigen was possible with the three antibodies tested. The polyclonal antibody was superior to the monoclonal antibodies, demonstrating good results with the three antigen retrieval protocols. The highest intensity staining was obtained with the citrate buffer and heat. The immunohistochemistry technique demonstrated the presence of viral antigens in the cytoplasm of neurons, in form of aggregates or with round or oval shape. The antigens were found as single or multiples inclusion bodies in the neurons. Immunohistochemistry is a fast method that can be used in routine procedures in cases where rabies is suspected, especially when the brain is submitted to the laboratory as formalin-fixed fragments or when samples could not be immediately shipped. The technique is also useful for retrospective studies. |
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