Introduction

Fumonisins B1, B2 and B3 are the main members of mycotoxins produced by Fusarium moniliforme. Fumonisins appear to be widespread in corn, and have been detected in corn-based products including feed and human foods. The presence of fumonisins in foods could have implications for human health. The Fumonisin EIA is intended for the quantitative detection of Fumonisin levels in rice, corn and cornmeal.

Principle of Fumonisin Determination

The enzyme immunoassay for Fumonisin is based on the competition between the fumonisin to be assayed and the Fumonisin-Horseradish Peroxidase conjugate, for binding to rabbit antibody directed against fumonisin, coated onto microwells. The sample containing the fumonisin, and the Fumonisin-Horseradish Peroxidase conjugate, when added to the microtiter wells, compete for binding to a limiting number of antibody sites. After incubation, each well is rinsed in order to remove non-bound components. The bound enzymatic activity is then measured by the addition of a chromogenic substrate. The intensity of the color developed is inversely proportional to the concentration of fumonisin in the sample. The concentration is calculated on the basis of a standard curve.

Reagents

• 96-well microtiter plate. Twelve strips of 8 detachable wells, coated with anti-Fumonisin antibody.
• One vial of 1.2 ml Negative Calibrator containing 0 μg/ml of fumonisin.
• Three vials of Positive Calibrator (0.9 ml) containing 1.0, 5.0 and 9.0 μg/ml of fumonisin.
• One bottle containing 10.5 ml of Fumonisin- Horseradish Peroxidase (FNS-HRP) Conjugate.
• One bottle containing 10.5 ml of Tetramethylbenzidine (TMB) substrate
• One bottle containing 15 ml of 10xWash Buffer (PBS-Tween).
• One bottle containing 10.5 ml of Stop Solution, 3N HCl

Assay Procedure

• Add 50 μl of standard or sample to the bottom of each well.
• Add 100 μl of FNS-HRP conjugate to each well. Incubate at room temperature for 30 minutes.
• Wash microtiter wells 3 times with wash buffer to remove the non-bound conjugate.
• Add 100 μl of TMB substrate to each well and incubate at room temperature for 15 min.
• Add 100 μl of stop solution to each well.
• Read absorbance at 450 nm using an ELISA reader.

Cross Reactivity

Do not cross react with aflatoxin, orchratoxin, zearalenone, deoxynivalenol and T2 toxin.

Fumonisin Inhibition Curve