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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Online Access: | http://dx.doi.org/10.1155/2013/593219 |
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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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