Development of a cell-based functional assay for the detection of clostridium botulinum neurotoxin types A and e

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

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Article number593219
JournalInternational Journal of Microbiology
Volume2013
DOIs
StatePublished - Apr 3 2013

Fingerprint

Clostridium botulinum
Type A Botulinum Toxins
Biological Assay
Clostridium
Neurotoxins
Laboratory Animals
Fluorescence
Food

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Microbiology (medical)

Cite this

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abstract = "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.",
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Development of a cell-based functional assay for the detection of clostridium botulinum neurotoxin types A and e. / Basavanna, Uma; Muruvanda, Tim; Brown, Eric Wayne; Sharma, Shashi K.

In: International Journal of Microbiology, Vol. 2013, 593219, 03.04.2013.

Research output: Contribution to journalArticle

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