Development of a Cell-Based Functional Assay for the Detection of Clostridium botulinum Neurotoxin Types A and E
| Autor: | Uma Basavanna, Tim Muruvanda, Eric W. Brown, Shashi K. Sharma |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: | |
| Zdroj: | International Journal of Microbiology, Vol 2013 (2013) |
| Druh dokumentu: | article |
| ISSN: | 1687-918X 1687-9198 |
| DOI: | 10.1155/2013/593219 |
| Popis: | 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. |
| Databáze: | Directory of Open Access Journals |
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