Produção de estoques virais centrifugados e ultracentrifugados do denv-2 para comparação dos títulos virais

Abstract

Dengue arbovirus represents a major challenge for Public Health in many of its aspects. This research is justified by the importance of establishing an in vitro viral mass stock with C6/36 cells infected by DENV-2, so that further studies are possible by research groups, which test natural products from the Amazon. The objectives were: to induce the multiplication of DENV-2 in cell strain of the aedes albopictus mosquito, C6/36; compare the titration of each viral stock through Plaque Forming Units (PFU) in VERO cells (African green monkey kidney cells), after centrifugation and ultracentrifugation; and differentiate viral, centrifugated and ultracentrifugated titrations. The production of dengue viral masses was performed in C6/36 cells. For viral titration, VERO cells were seded into plates of 6 wells and infected with ultracentrifugated DENV-2 for subsequent count of viral colonies, using the PFU technique. However, the experiment did not show positivity, some factors may have influenced: agarosis did not solidify when the neutral red dye was added, the dye itself may have generated influence, the ultracentrifugation process by rotating at 45,000 g during 90min at 4 ºC may have damaged viral stock or lack of infection. Therefore, it was possible to induce the multiplication of cells in c6/36 mosquito strains, centrifuge of cells infected by DENV-2 and ultracentrifugation. It was not possible to analyze and compare the titration of each viral stock using the PFU technique, after centrifugation and ultracentrifugation. Further tests should be carried out in order to compare viral stocks, enabling possible studies that test natural products from the Amazon. Keywords: Arboviruses; Viral mass; Viral titration.