Zearalenone ELISA Test Kit

Product Code ZEA-0050
Kit contains approximately 42 test samples plus controls, contains 6 microtiter strips.

Zearalenone, also known as RAL and F-2 mycotoxin, is a potent estrogenic metabolite produced by some Giberella species, a fungal plant pathogen. Several Fusarium species produce toxic substances of considerable concern to livestock and poultry producers: namely, deoxynivalenol, T-2 toxin, HT-2 toxin, diacetoxyscirpenol (DAS) and zearalenone. Zearalenone is the primary toxin causing infertility, abortion or other breeding problems, especially in swine. Zearalenone is heat-stable and is found worldwide in a number of cereal crops, such as maize, barley, oats, wheat, rice, and sorghum and also in bread. Commonly used extraction solvents are aqueous mixtures of methanol, acetonitrile or ethyl acetate followed by a range of different clean-up procedures that depend in part on the food and on the detection method in use. TLC methods and HPLC are commonly used. HPLC alone is not sufficient as it may often yield false positive results.

PRINCIPLE OF ASSAY

The zearalenone ELISA is a solid phase direct competitive enzyme immunoassay. A zearalenone-specific antibody optimized to react with zearalenone is coated to a polystyrene microwell. Toxins are extracted from a ground sample with 70% methanol. If zearalenone is present it will bind to the coated antibody. Subsequently, zearalenone bound to horse-radish peroxidase (HRP) is added and binds to the antibody not already occupied by zearalenone present in the sample or standard. After this incubation period, the contents of the wells are decanted, washed and an HRP substrate is added which develops a blue color in the presence of enzyme. The intensity of the color is directly proportional to the amount of bound conjugate and inversely proportional to the amount of zearalenone in the standard or sample. Therefore, as the concentration of zearalenone in the sample or standard increases, the intensity of the blue color will decrease. An acidic stop solution is added which changes the chromogen color from blue to yellow. The microwells are measured optically by a microplate reader with an absorbance filter of 450nm (OD450). The optical densities of the samples are compared to the OD’s of the kit standards and a result is determined by interpolation from the standard curve.