ABSTRACTObjectiveThis study verified the replication efficiency of the Rocio virus in a primary culture of mouse neural cells.MethodsMixed primary cultures (neurons/glia) obtained from the brains of newborn isogenic BALB/c mice were inoculated with Rocio virus on the 7thday of culture, and the development of cytopathogenic effects was monitored. The infection was confirmed via immunocytochemistry (anti-ROCV), while viral replication was quantified in infected primary cultures. The titration method used depended on the infection period.ResultsRocio virus efficiently infected primary cultured neural cells, with the highest viral titer causing cytopathic changes was observed at 2 days post infection. The virus-infected primary culture survived for up to 7 days post infection, and viral load quantitation showed viral replication kinetics compatible with the cell death kinetics of cultures.ConclusionThe findings of this study suggest that mouse neural cell primary cultures support Rocio virus replication and could be used as an alternative system for studyingFlavivirusinfection in the central nervous system.