Qualitative and Quantitative Detection of Botulinum Neurotoxins from Complex Matrices: Results of the First International Proficiency Test
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 appl...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2015-11-01
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| Series: | Toxins |
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| Online Access: | http://www.mdpi.com/2072-6651/7/12/4857 |
| _version_ | 1828369833885958144 |
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| author | Sylvia Worbs Uwe Fiebig Reinhard Zeleny Heinz Schimmel Andreas Rummel Werner Luginbühl Brigitte G. Dorner |
| author_facet | Sylvia Worbs Uwe Fiebig Reinhard Zeleny Heinz Schimmel Andreas Rummel Werner Luginbühl Brigitte G. Dorner |
| author_sort | Sylvia Worbs |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-14T06:29:55Z |
| format | Article |
| id | doaj.art-72c038b10daf43f889b61977d538d62a |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-14T06:29:55Z |
| publishDate | 2015-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-72c038b10daf43f889b61977d538d62a2022-12-22T02:07:39ZengMDPI AGToxins2072-66512015-11-017124935496610.3390/toxins7124857toxins7124857Qualitative and Quantitative Detection of Botulinum Neurotoxins from Complex Matrices: Results of the First International Proficiency TestSylvia Worbs0Uwe Fiebig1Reinhard Zeleny2Heinz Schimmel3Andreas Rummel4Werner Luginbühl5Brigitte G. Dorner6Biological Toxins, Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Seestrasse 10, 13353 Berlin, GermanyBiological Toxins, Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Seestrasse 10, 13353 Berlin, GermanyEuropean Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Retieseweg 111, 2440 Geel, BelgiumEuropean Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Retieseweg 111, 2440 Geel, Belgiumtoxogen GmbH, Feodor-Lynen-Strasse 35, 30625 Hannover, GermanyChemStat, Aarstrasse 98, 3005 Bern, SwitzerlandBiological Toxins, Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Seestrasse 10, 13353 Berlin, GermanyIn 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.http://www.mdpi.com/2072-6651/7/12/4857proficiency testBoNTreference materialstandardized detection |
| spellingShingle | Sylvia Worbs Uwe Fiebig Reinhard Zeleny Heinz Schimmel Andreas Rummel Werner Luginbühl Brigitte G. Dorner Qualitative and Quantitative Detection of Botulinum Neurotoxins from Complex Matrices: Results of the First International Proficiency Test Toxins proficiency test BoNT reference material standardized detection |
| title | Qualitative and Quantitative Detection of Botulinum Neurotoxins from Complex Matrices: Results of the First International Proficiency Test |
| title_full | Qualitative and Quantitative Detection of Botulinum Neurotoxins from Complex Matrices: Results of the First International Proficiency Test |
| title_fullStr | Qualitative and Quantitative Detection of Botulinum Neurotoxins from Complex Matrices: Results of the First International Proficiency Test |
| title_full_unstemmed | Qualitative and Quantitative Detection of Botulinum Neurotoxins from Complex Matrices: Results of the First International Proficiency Test |
| title_short | Qualitative and Quantitative Detection of Botulinum Neurotoxins from Complex Matrices: Results of the First International Proficiency Test |
| title_sort | qualitative and quantitative detection of botulinum neurotoxins from complex matrices results of the first international proficiency test |
| topic | proficiency test BoNT reference material standardized detection |
| url | http://www.mdpi.com/2072-6651/7/12/4857 |
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