A rapid and accurate assay for assessing the cytotoxicity of viral proteins.
A fluorescence-based assay is presented for measuring the cytoxicity of viral proteins added exogenously to cells. The assay is based on the use of two fluorescent dyes, calcein-AM and ethidium homodimer (EtD-1) to specifically stain living and dead cells respectively and employs fluorescence activated cells sorting (FACS) to achieve a rapid and accurate measurement of the cytotoxic capacity of a potential viral toxin. The assay has been developed using the group B homologue (ADRV-NSP4) of the NSP4 enterotoxin encoded by Group A rotaviruses but should be applicable to assaying any viral protein exhibiting cytotoxic activity.
Guzman
E
E
Department of Biological Sciences, University of Warwick, Gibbet Hill Rd., Coventry CV4 7AL, UK.
McCrae
M A
MA
D003601:Cytotoxicity Tests, Immunologic; D005456:Fluorescent Dyes; D006023:Glycoproteins / Q000633:toxicity; D018675:Toxicity Tests / Q000379:methods*; D014118:Toxins, Biological / Q000633:toxicity; D017361:Viral Nonstructural Proteins / Q000633:toxicity; D014764:Viral Proteins / Q000633:toxicity*
D005456:Fluorescent Dyes; D006023:Glycoproteins; C081369:NS28 protein, rotavirus; D014118:Toxins, Biological; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins