Recommended Immunological Strategies to Screen for Botulinum Neurotoxin-Containing Samples

Toxins

Uwe Fiebig¹  Martin B. Dorner¹  Sylvia Worbs¹  Tanja Endermann¹  Stéphanie Simon²  Hervé Volland²  Marie-Claire Nevers²  Julie Dano²  Dorothea Sesardic³  Rob Tierney³  Yvonne Liu³ 

[1] Biological Toxins, Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Seestrasse 10, 13353 Berlin, Germany;CEA Saclay, Institute of Biology and Technologies of Saclay, Laboratory for Immunoanalytical Researches, Gif-sur-Yvette 91191 cedex, France;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;

关键词: proficiency test;    botulinum neurotoxin;    Clostridium botulinum;    immunological detection;    ELISA;    lateral flow immunoassay;    endopeptidase;    antibodies;    matrices;   

DOI  :  10.3390/toxins7124860

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown