Growth of Clostridium (C.) botulinum in foods results in the production of botulinum neurotoxin (BoNT), which, when ingested, leads to foodborne botulism. C. botulinum spores are widely distributed in foods and the environment. Detection of C. botulinum requires enrichment culture of samples followed by screening of these cultures for the presence of BoNT or polymerase chain reaction detection of genes encoding BoNT. While BoNT is traditionally detected using a mouse bioassay, several types of in vitro assays have been developed using combinations of immunoassays, endopeptidase, and cell-based assays.