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Toxins 2017, 9(4), 145; doi:10.3390/toxins9040145

A Simple and Specific Noncompetitive ELISA Method for HT-2 Toxin Detection

1
VTT Technical Research Centre of Finland, Tietotie 2 FI-02150 Espoo, Finland
2
Finnish Food Safety Authority (Evira), Chemistry and Toxicology Unit, Research and Laboratory Department, Mustialankatu 3, FI-00790 Helsinki, Finland
*
Author to whom correspondence should be addressed.
Academic Editors: Michelangelo Pascale and Maria C. DeRosa
Received: 24 March 2017 / Revised: 13 April 2017 / Accepted: 14 April 2017 / Published: 20 April 2017
(This article belongs to the Collection Biorecognition Assays for Mycotoxins)
View Full-Text   |   Download PDF [2069 KB, uploaded 20 April 2017]   |  

Abstract

We developed an HT-2 toxin-specific simple ELISA format with a positive read-out. The assay is based on an anti-immune complex (IC) scFv antibody fragment, which is genetically fused with alkaline phosphatase (AP). The anti-IC antibody specifically recognizes the IC between a primary anti-HT-2 toxin Fab fragment and an HT-2 toxin molecule. In the IC ELISA format, the sample is added together with the scFv-AP antibody to the ELISA plate coated with the primary antibody. After 15 min of incubation and a washing step, the ELISA response is read. A competitive ELISA including only the primary antibody recognizes both HT-2 and T-2 toxins. The anti-IC antibody makes the assay specific for HT-2 toxin, and the IC ELISA is over 10 times more sensitive compared to the competitive assay. Three different naturally contaminated matrices: wheat, barley and oats, were used to evaluate the assay performance with real samples. The corresponding limits of detection were 0.3 ng/mL (13 µg/kg), 0.1 ng/mL (4 µg/kg) and 0.3 ng/mL (16 µg/kg), respectively. The IC ELISA can be used for screening HT-2 toxin specifically and in relevant concentration ranges from all three tested grain matrices. View Full-Text
Keywords: HT-2 toxin; recombinant antibodies; ELISA; noncompetitive; Fab; scFv; alkaline phosphatase; fusion protein; cereal grains HT-2 toxin; recombinant antibodies; ELISA; noncompetitive; Fab; scFv; alkaline phosphatase; fusion protein; cereal grains
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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

Arola, H.O.; Tullila, A.; Nathanail, A.V.; Nevanen, T.K. A Simple and Specific Noncompetitive ELISA Method for HT-2 Toxin Detection. Toxins 2017, 9, 145.

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