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

Appl Environ Microbiol. 1996 Jun;62(6):1880-4. doi: 10.1128/aem.62.6.1880-1884.1996.

Abstract

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 (H202-3,3',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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Evaluation Studies as Topic
  • Fusarium / chemistry*
  • Fusarium / immunology
  • Fusarium / isolation & purification
  • Immunoenzyme Techniques*
  • Membranes, Artificial
  • Mice
  • Rabbits
  • T-2 Toxin / analysis*
  • T-2 Toxin / immunology
  • Triticum / chemistry*
  • Triticum / microbiology*

Substances

  • Antibodies, Monoclonal
  • Membranes, Artificial
  • T-2 Toxin