Dipstick enzyme immunoassay to detect Fusarium T-2 toxin in wheat

A dipstick enzyme immunoassay for the rapid detection of Fusarium T-2 toxin in wheat was developed, An Immunodyne ABC membrane was precoated with rabbit anti mouse immunoglobulins. After the strips were immersed in a solution of monoclonal anti-T-2 toxin antibodies, a direct competitive enzyme immunoassay was performed. This assay included the incubation of the antibody-coated dipsticks in a mixture of sample and T-2 toxin-horseradish peroxidase conjugate. Afterwards, the strips were placed in a chromogen-containing substrate solution (H2O2-3,3',5,5'-tetramethylbenzidine) for color reaction, The dot color intensity of toxin-positive dipsticks was visually distinguishable from that of the negative control, A portable colorimeter was used to confirm and quantify the visual observations. With coated strips, the tests could be performed in 45 min, The visual detection limit for T-2 toxin in buffer solution was 0.25 ng/ml. Artificially infected wheat samples were extracted with 80% methanol-water. A dilution of the raw extract of 1:8 was sufficient to avoid matrix effects. It was possible to make visually a clear distinction between the negative control and a wheat extract spiked with 12 ng/g.