Unknown

Dataset Information

0

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


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.

SUBMITTER: Arola HO 

PROVIDER: S-EPMC5408219 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature

altmetric image

Publications

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

Arola Henri O HO   Tullila Antti A   Nathanail Alexis V AV   Nevanen Tarja K TK  

Toxins 20170420 4


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  ...[more]

Similar Datasets

| S-EPMC2793542 | biostudies-literature
| S-EPMC6221316 | biostudies-literature
| S-EPMC8066203 | biostudies-literature
| S-EPMC3315793 | biostudies-literature
| S-EPMC7655971 | biostudies-literature
| S-EPMC5198558 | biostudies-literature
| S-EPMC7057676 | biostudies-literature
| S-EPMC6225493 | biostudies-literature
| S-EPMC10534902 | biostudies-literature
| S-EPMC6821646 | biostudies-literature