title: Development of a Cell-Based Functional Assay for the Detection of Clostridium botulinum Neurotoxin Types A and E.
abstract: The standard procedure for definitive detection of BoNT-producing Clostridia is a culture method combined with neurotoxin detection using a standard mouse bioassay (MBA). The mouse bioassay is highly sensitive and specific, but it is expensive and time-consuming, and there are ethical concerns due to use of laboratory animals. Cell-based assays provide an alternative to the MBA in screening for BoNT-producing Clostridia. Here, we describe a cell-based assay utilizing a fluorescence reporter construct expressed in a neuronal cell model to study toxin activity in situ. Our data indicates that the assay can detect as little as 100 pM BoNT/A activity within living cells, and the assay is currently being evaluated for the analysis of BoNT in food matrices. Among available in vitro assays, we believe that cell-based assays are widely applicable in high-throughput screenings and have the potential to at least reduce and refine animal assays if not replace it.
authors.affiliation: Division of Microbiology, Office of Regulatory Science, Center for Food Safety and Applied Nutrition, Food and Drug Administration, CPK1, HFS-712, 5100 Paint Branch Parkway, College Park, MD 20740, USA.
authors.forename: Uma
authors.initials: U
authors.lastname: Basavanna
authors.forename: Tim
authors.initials: T
authors.lastname: Muruvanda
authors.forename: Eric W
authors.initials: EW
authors.lastname: Brown
authors.forename: Shashi K
authors.initials: SK
authors.lastname: Sharma