Qualitative and Quantitative Detection of Botulinum Neurotoxins from Complex Matrices: Results of the First International Proficiency Test

Toxins

Sylvia Worbs¹  Uwe Fiebig¹  Reinhard Zeleny⁴  Heinz Schimmel⁴  Andreas Rummel²  Werner Luginbühl³  Brigitte G. Dorner¹ 

[1] Biological Toxins, Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Seestrasse 10, 13353 Berlin, Germany;toxogen GmbH, Feodor-Lynen-Strasse 35, 30625 Hannover, Germany;ChemStat, Aarstrasse 98, 3005 Bern, Switzerland;European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Retieseweg 111, 2440 Geel, Belgium;

关键词: proficiency test;    BoNT;    reference material;    standardized detection;   

DOI  :  10.3390/toxins7124857

来源: mdpi

【 摘 要 】

In the framework of the EU project EQuATox, a first international proficiency test (PT) on the detection and quantification of botulinum neurotoxins (BoNT) was conducted. Sample materials included BoNT serotypes A, B and E spiked into buffer, milk, meat extract and serum. Different methods were applied by the participants combining different principles of detection, identification and quantification. Based on qualitative assays, 95% of all results reported were correct. Successful strategies for BoNT detection were based on a combination of complementary immunological, MS-based and functional methods or on suitable functional in vivo/in vitro approaches (mouse bioassay, hemidiaphragm assay and Endopep-MS assay). Quantification of BoNT/A, BoNT/B and BoNT/E was performed by 48% of participating laboratories. It turned out that precise quantification of BoNT was difficult, resulting in a substantial scatter of quantitative data. This was especially true for results obtained by the mouse bioassay which is currently considered as “gold standard” for BoNT detection. The results clearly demonstrate the urgent need for certified BoNT reference materials and the development of methods replacing animal testing. In this context, the BoNT PT provided the valuable information that both the Endopep-MS assay and the hemidiaphragm assay delivered quantitative results superior to the mouse bioassay.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.