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Issue 20, 2013
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Bioluminescence assay for the highly sensitive detection of botulinum neurotoxin A activity

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Abstract

This article describes a novel bioluminescence assay for detecting the proteolytic activity of Botulinum NeuroToxins (BoNT) in complex matrices. The assay is capable of detecting traces of BoNT in blood samples as well as in food drinks. The assay was responsive to BoNT/A subtypes 1 to 5, and serotype E3 in buffered solutions. It was responsive to filtered Clostridium botulinum supernatants and BoNT/A1 in complex with neurotoxin associated proteins in bouillon and milk (3.8% fat) down to 400 fM after 4 h RT incubation and in bouillon at concentrations down to 120 fM after 21 h RT incubation. In combination with an immunocapture/enrichment step it could detect BoNT/A1 in citrated plasma at concentrations down to 30 fM (1.2 mouse LD50 per mL). The simplicity of the assay, combined with a demonstrated ability to lyophilize the reagents, demonstrates its usefulness for detection of BoNT in non-specialised analytical laboratories.

Graphical abstract: Bioluminescence assay for the highly sensitive detection of botulinum neurotoxin A activity

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Supplementary files

Article information


Submitted
15 mar. 2013
Accepted
29 júl. 2013
First published
31 júl. 2013

Analyst, 2013,138, 6154-6162
Article type
Paper

Bioluminescence assay for the highly sensitive detection of botulinum neurotoxin A activity

G. B. Stevens, D. A. Silver, A. Zgaga-Griesz, W. G. Bessler, S. K. Vashist, P. Patel, K. Achazi, J. Strotmeier, S. Worbs, M. B. Dorner, B. G. Dorner, D. Pauly, A. Rummel, G. A. Urban and M. Krueger, Analyst, 2013, 138, 6154
DOI: 10.1039/C3AN00525A

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