Production and characterization of Chikungunya virus for strategic antigen development to support public health demands

Abstract

Methodology: Vero cells infection by CHIKV was conducted with a Multiplicity of Infection (MOI) of 1.0. Monitoring virus entry, cells were fixed, embedded in polymers, ultrathin cut and analyzed by transmission electron microscopy (TEM). Quantification of CHIKV was performed by real-time PCR (qPCR), for the particles inside the cell and in the supernatant, and by plaque assay for supernatant. Investigating the best conditions for CHIKV production, kinetics were conducted in two MOIs (0.01 and 0.005) for 48 h. Production in serum-free media was also tested. Kinetics were monitored by cytopathic effect (CPE), plaque assay and qPCR.