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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| Format: | Article |
| Language: | English |
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Wiley
2013-01-01
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| Series: | International Journal of Microbiology |
| Online Access: | http://dx.doi.org/10.1155/2013/593219 |
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| author | Uma Basavanna Tim Muruvanda Eric W. Brown Shashi K. Sharma |
| author_facet | Uma Basavanna Tim Muruvanda Eric W. Brown Shashi K. Sharma |
| author_sort | Uma Basavanna |
| collection | DOAJ |
| 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. |
| format | Article |
| id | doaj-art-69b4f9f7dd534ae186a509c00da99070 |
| institution | Kabale University |
| issn | 1687-918X 1687-9198 |
| language | English |
| publishDate | 2013-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | International Journal of Microbiology |
| spelling | doaj-art-69b4f9f7dd534ae186a509c00da990702025-02-03T06:45:56ZengWileyInternational 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 |
| 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 |
| title | 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_short | 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 |
| url | http://dx.doi.org/10.1155/2013/593219 |
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