Plaque Assay to Evaluate Antiviral Effects of a Plant-Derived Compound on Zika Virus

Take a multi-well plate containing epithelial cells.

Treat one well with Zika virus alone as the control, and another well with Zika virus plus silvestrol, a plant-derived antiviral compound, as the test.

Incubate to allow viral adsorption onto the cells.

Remove the medium and wash the wells to remove unbound viruses.

Overlay the wells with a medium containing agarose to restrict viral diffusion and incubate.

In the control, the virus is endocytosed into the cell.

It directs the synthesis of viral RNA and proteins, forming new viral particles that cause cell lysis and spread locally, creating plaques.

In the test, silvestrol interferes with viral protein synthesis, limiting viral production and resulting in fewer plaques.

Remove the agarose and treat the cells with a fixative.

Add a dye to stain the cells, while the plaques remain clear.

Fewer plaques appear in the silvestrol-treated well than in the control, confirming its antiviral activity.