A Simple and Specific Noncompetitive ELISA Method for HT-2 Toxin Detection
AbstractWe 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
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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.
Arola HO, Tullila A, Nathanail AV, Nevanen TK. A Simple and Specific Noncompetitive ELISA Method for HT-2 Toxin Detection. Toxins. 2017; 9(4):145.Chicago/Turabian Style
Arola, Henri O.; Tullila, Antti; Nathanail, Alexis V.; Nevanen, Tarja K. 2017. "A Simple and Specific Noncompetitive ELISA Method for HT-2 Toxin Detection." Toxins 9, no. 4: 145.
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