Development of a Cell-Based Functional Assay for the Detection of Clostridium botulinum Neurotoxin Types A and E

Uma Basavanna; Tim Muruvanda; Eric W. Brown; Shashi K. Sharma

doi:10.1155/2013/593219

International Journal of Microbiology (Jan 2013)

Development of a Cell-Based Functional Assay for the Detection of Clostridium botulinum Neurotoxin Types A and E

Uma Basavanna,
Tim Muruvanda,
Eric W. Brown,
Shashi K. Sharma

Affiliations

Uma Basavanna: 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
Tim Muruvanda: 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
Eric W. Brown: 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
Shashi K. Sharma: 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

DOI: https://doi.org/10.1155/2013/593219
Journal volume & issue: Vol. 2013

Abstract

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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.

Published in International Journal of Microbiology

ISSN: 1687-918X (Print); 1687-9198 (Online)
Publisher: Hindawi Limited
Country of publisher: United Kingdom
LCC subjects: Science: Microbiology
Website: https://onlinelibrary.wiley.com/journal/8472

About the journal