DEVELOPMENT AND STANDARDIZATION OF AN INDIRECT ELISA FOR THE DIAGNOSIS MAEDI-VISNA IN SHEEP
DOI:
https://doi.org/10.5216/cab.v9i1.3677Keywords:
Medicina Veterinária PreventivaAbstract
The objective of this work was to develop and standardize an indirect ELISA for diagnosis of Maedi Visna (MV) infection. The antigen was produced from the supernatants of caprine synovial membrane (CSM) cell monolayers inoculated with Maedi Visna virus (MVV), strain K1514, by cycles of freezing and thawed, and clarified by centrifugation at 3000 g for 40 min. The clarified suspension was precipitated with PEG 8000 and centrifugated at 12000 g for 60 min; the pellet was resuspended in buffer TNE and layered onto a sucrose cushion by centrifugation at 42000 g for 105 min. The pellet was resuspended in PBS containing phenylmethylsulphonyl fluoride (PMSF). The ELISA was performed in 96 wells plates, incubated for 1 h at 37 ºC. The reaction was detected by incubation with the enzyme substrate and o-phenylenediamine (OPD) during 15 min. The tests were compared using 175 sera samples. The concentration of the antigen was 2 µg/mL and the best dilution of the sera was 1:100. ELISA detected more positive samples (22.9%) than AGID (6.3%) and presented a better sensitivity than AGID and, although the specificity was below the expected, it could already be recomended in the diagnosis of MVV infection.KEY WORDS: ELISA, MaediVisna, serologic diagnosis, sheep lentivirus.Downloads
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Published
2008-04-03
How to Cite
DANTAS, T. V. M.; ARAÚJO, S. A. C. de; PINHEIRO, R. R.; ARAGÃO, M. A. do C.; SILVA, J. B. A. da; RICARTE, A. R. F.; RIBEIRO, A. L.; TEIXEIRA, M. F. da S. DEVELOPMENT AND STANDARDIZATION OF AN INDIRECT ELISA FOR THE DIAGNOSIS MAEDI-VISNA IN SHEEP. Brazilian Animal Science/ Ciência Animal Brasileira, Goiânia, v. 9, n. 1, p. 181–187, 2008. DOI: 10.5216/cab.v9i1.3677. Disponível em: https://revistas.ufg.br/vet/article/view/3677. Acesso em: 8 nov. 2024.
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Veterinary Medicine
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