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T-2 Toxin/HT-2 Toxin and Ochratoxin A ELISAs Development and In-House Validation in Food in Accordance with Commission Regulation (EU) No 519/2014

EuroProxima B.V., Arnhem 6827 BN, The Netherlands
RIKILT Wageningen UR, Wageningen 6708 WB, The Netherlands
Institute for Global Food Security, School of Biological Sciences, Queen’s University Belfast, Belfast BT9 5BN, UK
Author to whom correspondence should be addressed.
Academic Editors: Michelangelo Pascale and Maria C. DeRosa
Toxins 2017, 9(12), 388;
Received: 15 November 2017 / Revised: 25 November 2017 / Accepted: 29 November 2017 / Published: 30 November 2017
(This article belongs to the Collection Biorecognition Assays for Mycotoxins)
PDF [712 KB, uploaded 16 December 2017]


T-2 toxin/HT-2 toxin (T-2/HT-2) and ochratoxin A (OTA) are mycotoxins that can contaminate a variety of agricultural commodities. To protect consumers’ health, indicative limits for T-2/HT-2 and maximum limits for OTA have been set by the European Commission, requiring food business operators and controlling agencies to conduct routine checks for the presence of these harmful contaminants. Screening methods are increasingly used for monitoring purposes. Due to the demand for new and improved screening tools, two individual detection methods, T-2/HT-2 and OTA enzyme-linked immunosorbent assays (ELISAs), were developed in this study. The T-2/HT-2 ELISA was based on a T-2 monoclonal antibody with an IC50 (50% inhibitory concentration) of 0.28 ng/mL and 125% cross-reactivity with HT-2. As regards the OTA ELISA, a new sensitive monoclonal antibody specific to OTA with an IC50 of 0.13 ng/mL was produced. Both developed ELISA tests were then validated in agricultural commodities in accordance with the new performance criteria guidelines for the validation of screening methods for mycotoxins included in Commission Regulation (EU) No 519/2014. The T-2/HT-2 ELISA was demonstrated to be suitable for the detection of T-2/HT-2 in cereals and baby food at and above the screening target concentration (STC) of 12.5 μg/kg and 7.5 μg/kg, respectively. The OTA ELISA was shown to be applicable for the detection of OTA in cereals, coffee, cocoa and wine at and above the STC of 2 μg/kg, 2.5 μg/kg, 2.5 μg/kg and 0.4 ng/mL, respectively. The accuracy of both ELISAs was further confirmed by analysing proficiency test and reference samples. The developed methods can be used for sensitive and high-throughput screening for the presence of T-2/HT-2 and OTA in agricultural commodities. View Full-Text
Keywords: mycotoxins; enzyme-linked immunosorbent assay; immunoassay; screening mycotoxins; enzyme-linked immunosorbent assay; immunoassay; screening

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Oplatowska-Stachowiak, M.; Kleintjens, T.; Sajic, N.; Haasnoot, W.; Campbell, K.; Elliott, C.T.; Salden, M. T-2 Toxin/HT-2 Toxin and Ochratoxin A ELISAs Development and In-House Validation in Food in Accordance with Commission Regulation (EU) No 519/2014. Toxins 2017, 9, 388.

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