ENAGIS-MAY: uma proposta de ensaio de aglutinação indireta para detectar no sangue imunoglobulinas contra o vírus Mayaro

• Autor(a) principal: Oliveira, Marcelo Santos de
• Data de Publicação: 2008
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: LOCUS Repositório Institucional da UFV
• Texto Completo: http://locus.ufv.br/handle/123456789/2316
• topic: Mayaro; Aglutinação passiva; Diagnóstico; Mayaro; Passive agglutination; Diagnostic; CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL

Mayaro virus (MAYV) was initially isolated in Trinidad in 1954. In Brazil was isolated in 1955 during an outbreak in the state of Pará. Various outbreaks have been described in areas of the Amazon forest, in the north of Brazil, where it can be considered endemic, and whose ecosystem has been deeply disturbed. The symptoms caused by MAYV infection are similar to the caused by several arboviral diseases, include dengue. The objective of this work was the establishment of an indirect agglutination assay for the detection of immunoglobulins against Mayaro virus using whole blood (ENAGIS- MAY). VERO cells were infected with MAYV (MOI 1) to obtaining antigens, which were covalently coupled to carboxyl-modified polystyrene beads. For the biological assays eight Mus muscullus, Swiss lineage mice were used, divided in to two groups. Group M was infected with MAYV and group C received PBS pH 7,2. After 30 days, blood samples were collected and analyzed for the presence of antimayaro antibodies. About 50 &#956;L of blood from each animal were applied to filter paper disks and, after drying, eluated with 0,2M borate buffer pH 8,5. ENAGIS-MAY was assayed with 10&#956;L of eluate and 10&#956;L of suspension of latex beads containing virus proteins. After 5 min incubation, the occurrence of agglutination was observed. Samples in group M presented titers of 50, 100, 50 and 50 for M1, M2, M3 and M4, respectively, while all samples in group C were negative. Titers of neutralizing antibodies were determined for the two groups by the microculture plaque neutralization test and used as gold standard to calculate the sensibility (100%; 95% C.I.: 39,8- 100,0) and specificity (100%; 95% C.I.: 9,8-100,0) of ENAGIS- MAY. Analysis by the Fisher test indicated that the assays results were significantly correlated (p <0,05). Therefore ENAGISMAY is likely to be feasible for routine diagnoses of the Mayaro fever using blood-impregnated filter paper.