A Cell Membrane Fusion Assay for the Fish Pathogen Spring Viremia of Carp Virus (SVCV).

Abstract

Fusion of the infected cell membranes is a characteristic effect of a wide number of viral infections. Spring viremia of carp virus (SVCV), a rhabdovirus causing disease in farmed carp, can induce membrane fusion of the infected cells by shifting the pH of the culture medium to slightly acidic. Membrane fusion leads to the formation of clusters of cell nuclei enclosed in a cell membrane, the so-called syncytia, that can be easily visualized by cell staining and light microscope inspection. In the present work, we report a protocol to induce syncytia formation in EPC cells infected with SVCV, where membrane fusion is triggered by a low-pH incubation step. Appearance of syncytia can be observed at 18 hours post infection. The syncytia formation assay described here may serve as an experimental platform to quantitate SVCV, to determine virus infectivity, and a useful tool to study virus entry into the cell as well as to test candidate antiviral compounds that could block the entry of SVCV into cells by inhibiting membrane fusion.