Padronização do Elisa indireto e Western Blot para diagnóstico da artrite-encefalite caprina

• Autor(a) principal: Rodrigues, A.s.
• Data de Publicação: 2014
• Outros Autores: Brito, R.l.l. [UNESP], Pinheiro, R.r., Dias, R.p., Alves, S.m., Souza, T.s., Souza, K.c., Azevedo, D.a.a., Andrioli, A., Magalhães, D.c.t., Teixeira, M.f.s.
• Tipo de documento: Artigo
• Idioma: por
• Título da fonte: Repositório Institucional da UNESP
• Texto Completo: http://dx.doi.org/10.1590/1678-41626303; http://hdl.handle.net/11449/109524
• topic: especificidade; infecção; LVPR; sensibilidade; sorologia; infection; sensitivity; serology; specificity; SRLV

Caprine arthritis-encephalitis (CAE) is routinely diagnosed with the Agarose Gel Immunodiffusion (AGID) technique, which is considered to have low sensitivity. The objective of this study was to standardize testing i-Elisa and Western Blot for early detection of antibodies against CAEV in goats and compare the results obtained in these tests with proof of AGID. For standardization of i-Elisa and WB, different concentrations and dilutions of antigen, sera and conjugate were used. In the i-Elisa, rigid microplate with 96 wells was adopted, and the combination that showed the best result was a concentration of 0.5µg/ well of antigen and dilutions of the serum of 1:100 and conjugate of 1:1500. In the WB nitrocellulose membranes were used, and the dilutions of the serum were defined at 1:50 and conjugate at 1:15000. To evaluate the performance of the techniques, 222 goat serum samples were tested and the data were compared with the AGID. The sensitivity and specificity of Elisa-i/IDGA, WB/AGID and WB/Elisa-i were 70% and 91%, 100% and 72.6%, 84.6% and 76.5%, concomitantly. The Kappa index of these tests was 0.35, 0.2 and 0.36, respectively. The i-Elisa and WB techniques were more sensitive than the AGID and can be used as tools for early diagnosis of CAE.