O vírus Vaccínia (VACV) é responsável por causar doença exantemática e vesicular em bovinos e também em seres humanos. As infecções causadas por estes vírus vêm sendo frequentemente relatadas nos últimos 10 anos no Brasil. O objetivo deste trabalho foi monitorar os intervalos de replicação e montagem do VACV-WR (Western Reserve) em células Vero, com a finalidade de expor os principais meios de bloqueio do ciclo replicativo que podem ser utilizados por drogas antivirais. Células Vero foram infectadas com m.o.i de 0,1 da cepa VACV-WR. Posteriormente, as células foram fixadas em intervalos de tempo de 6, 12, 18, 24, 30, 36, 42 e 48 horas pós – infecção, e processadas para microscopia eletrônica de transmissão. A multiplicação de VACV-WR em monocamada de células Vero produziu efeito citopático (fusão de membranas, sincícios e vacúolos) em aproximadamente 90% das células. Análises das micrografias eletrônicas permitiram evidenciar detalhadamente o efeito citopático, a dinâmica dos distúrbios metabólicos 4celulares e o ciclo replicativo viral até a formação da partícula viral extracelular. Assim, este trabalho pôde mostrar possíveis alvos terapêuticos para inibição da replicação de VACV- WR e novas abordagens no uso de fármacos com ação antiviral.
Human adenoviruses comprise an important group of etiologic agents that are responsible for various diseases in adults and children, such as respiratory, ocular, gastroenteric, and urinary infections. In immunocompromised and organ-transplanted individuals, these agents can cause generalized infections. Rapid diagnostic methods for detecting these infectious agents are not widely available. The aim of this work was to produce monoclonal and polyclonal anti-adenovirus antibodies to be used in a rapid diagnostic test for respiratory infections. Adenovirus hexons were satisfactorily purified by ultracentrifugation and chromatography. After virus purification, anti-hexon monoclonal antibodies were produced and characterized, following classical methods. Antibodies were specific for adenoviruses 2, 3, 5, and 41. The proposed immunochromatographic test was standardized using colloidal gold. The standardization of the rapid test was sufficient to detect adenovirus antigens (in nasopharyngeal lavage samples) with sensitivity of 100% and specificity of 85% when compared to direct immunofluorescence. The immunochromatographic assay prototype was sufficiently sensitive to detect B (3), C (2 and 5), and F (41) adenovirus samples. Although based on preliminary data, the test demonstrated the same performance as direct immunofluorescence, but with the advantage of being a point-of-care test. Further studies are still needed to confirm its effectiveness in clinical practice.
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