Dengue Virus (DENV) and ZIKA virus (ZIKV) Antiviral Services
Antiviral assays against Dengue virus are performed to determine the inhibitory activity of small-molecule compounds and neutralizing antibodies for vaccine design. In vitro replication and infectivity assays against DENV-2 (16681 and TSV01) are routinely performed in monolayers of Vero cells. Several readouts are used to monitor virus replication, including specific ELISA assays and flow cytometry determination of intracellular viral antigens (e.g. capsid protein). These assays allow rapid and sensitive detection of Dengue virus replication in human and mosquito cell lines. Antiviral assays against other DENV strains are under development. DENV strains for each of the four known serotypes are available for these assays.
Flow-cytometry based assay to determine inhibition of DENV replication or neutralization with antibodies. In the above example incubation with test antibody 1 results in 60% inhibition of DENV infection. In the figure below dose-response studies with 4 mAbs neutralizing DENV-2 are shown. The assay is optimized and validated with DENV serotypes 1-4.
Antiviral assays against ZIKA virus are performed using the Virus CPE (cytophatic effect) prevention assay. In this assay the virus-induced CPE is prevented by the presence of an antiviral agent. The cytopathic effects mediated by the virus are monitored using neutral red. Viable cells incorporate neutral red in their lysosomes. Uptake of neutral red relies on the ability of live cells to maintain a lower pH inside their lysosomes than in the cytoplasm, and this active process requires ATP. Once inside the lysosome, the neutral red dye becomes charged and is retained intracellularly. Our standard assay is performed with the PLCal_ZV strain using Vero (epithelial kidney) cells. Over a dozen different strains of ZIKA are available for use in antiviral assays.
For additional information about antidengue or anti-ZIKA services call us at (858) 232-7919 or contact us at antivirals@retrovirox.com
To find out about other antiviral assays offered at RetroVirox click on the following links:
Human Respiratory Syncytial Virus (HRSV)
HIV Pseudoviruses for Neutralization Assays
