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

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 t...

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Main Authors:	Uma Basavanna, Tim Muruvanda, Eric W. Brown, Shashi K. Sharma
Format:	Article
Language:	English
Published:	Hindawi Limited 2013-01-01
Series:	International Journal of Microbiology
Online Access:	http://dx.doi.org/10.1155/2013/593219

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spelling	doaj-69b4f9f7dd534ae186a509c00da990702021-07-02T05:51:12ZengHindawi LimitedInternational Journal of Microbiology1687-918X1687-91982013-01-01201310.1155/2013/593219593219Development of a Cell-Based Functional Assay for the Detection of Clostridium botulinum Neurotoxin Types A and EUma Basavanna0Tim Muruvanda1Eric W. Brown2Shashi K. Sharma3Division 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, USADivision 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, USADivision 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, USADivision 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, USAThe 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.http://dx.doi.org/10.1155/2013/593219
collection	DOAJ
language	English
format	Article
sources	DOAJ
author	Uma Basavanna Tim Muruvanda Eric W. Brown Shashi K. Sharma
spellingShingle	Uma Basavanna Tim Muruvanda Eric W. Brown Shashi K. Sharma Development of a Cell-Based Functional Assay for the Detection of Clostridium botulinum Neurotoxin Types A and E International Journal of Microbiology
author_facet	Uma Basavanna Tim Muruvanda Eric W. Brown Shashi K. Sharma
author_sort	Uma Basavanna
title	Development of a Cell-Based Functional Assay for the Detection of Clostridium botulinum Neurotoxin Types A and E
title_short	Development of a Cell-Based Functional Assay for the Detection of Clostridium botulinum Neurotoxin Types A and E
title_full	Development of a Cell-Based Functional Assay for the Detection of Clostridium botulinum Neurotoxin Types A and E
title_fullStr	Development of a Cell-Based Functional Assay for the Detection of Clostridium botulinum Neurotoxin Types A and E
title_full_unstemmed	Development of a Cell-Based Functional Assay for the Detection of Clostridium botulinum Neurotoxin Types A and E
title_sort	development of a cell-based functional assay for the detection of clostridium botulinum neurotoxin types a and e
publisher	Hindawi Limited
series	International Journal of Microbiology
issn	1687-918X 1687-9198
publishDate	2013-01-01
description	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.
url	http://dx.doi.org/10.1155/2013/593219
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Development of a Cell-Based Functional Assay for the Detection of Clostridium botulinum Neurotoxin Types A and E

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