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Toxins 2015, 7(12), 5011-5034; doi:10.3390/toxins7124860

Recommended Immunological Strategies to Screen for Botulinum Neurotoxin-Containing Samples

1
CEA Saclay, Institute of Biology and Technologies of Saclay, Laboratory for Immunoanalytical Researches, Gif-sur-Yvette 91191 cedex, France
2
Biological Toxins, Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Seestrasse 10, 13353 Berlin, Germany
3
Division of Bacteriology, National Institute for Biological Standards and Control, a Centre of Medicines & Healthcare Products Regulatory Agency, Blanche Lane, South Mimms, Potters Bar, Hertfordshire EN6 3QG, UK
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editors: Andreas Rummel and Brigitte G. Dorner
Received: 15 October 2015 / Revised: 3 November 2015 / Accepted: 4 November 2015 / Published: 26 November 2015
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Abstract

Botulinum neurotoxins (BoNTs) cause the life-threatening neurological illness botulism in humans and animals and are divided into seven serotypes (BoNT/A–G), of which serotypes A, B, E, and F cause the disease in humans. BoNTs are classified as “category A” bioterrorism threat agents and are relevant in the context of the Biological Weapons Convention. An international proficiency test (PT) was conducted to evaluate detection, quantification and discrimination capabilities of 23 expert laboratories from the health, food and security areas. Here we describe three immunological strategies that proved to be successful for the detection and quantification of BoNT/A, B, and E considering the restricted sample volume (1 mL) distributed. To analyze the samples qualitatively and quantitatively, the first strategy was based on sensitive immunoenzymatic and immunochromatographic assays for fast qualitative and quantitative analyses. In the second approach, a bead-based suspension array was used for screening followed by conventional ELISA for quantification. In the third approach, an ELISA plate format assay was used for serotype specific immunodetection of BoNT-cleaved substrates, detecting the activity of the light chain, rather than the toxin protein. The results provide guidance for further steps in quality assurance and highlight problems to address in the future. View Full-Text
Keywords: proficiency test; botulinum neurotoxin; Clostridium botulinum; immunological detection; ELISA; lateral flow immunoassay; endopeptidase; antibodies; matrices proficiency test; botulinum neurotoxin; Clostridium botulinum; immunological detection; ELISA; lateral flow immunoassay; endopeptidase; antibodies; matrices
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Simon, S.; Fiebig, U.; Liu, Y.; Tierney, R.; Dano, J.; Worbs, S.; Endermann, T.; Nevers, M.-C.; Volland, H.; Sesardic, D.; Dorner, M.B. Recommended Immunological Strategies to Screen for Botulinum Neurotoxin-Containing Samples. Toxins 2015, 7, 5011-5034.

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