2 FORMATS OF ENZYME-IMMUNOASSAY FOR 15-ACETYLDEOXYNIVALENOL APPLIED TO WHEAT

Polyclonal antibodies against 15-acetyldeoxynivalenol (15-AcDON) were raised in rabbits after immunization with a 3-hemiglutaryl-15-AcDON (15-AcDON-HG) derivative coupled to human serum albumin. Using these antibodies and a 15-AcDON-HG-horseradish peroxidase conjugate in direct enzyme immunoassay (EIA), a microtiter plate EIA and a membrane-based dipstick EIA were established, with detection limits for 15-AcDON of 0.35 and 5 ng/mL, respectively. When specificity was checked on the basis of the 50% inhibition levels, relative cross-reactivities with 15-AcDON, diacetyl-DON, triacetyl-DON, 3-AcDON, and DON were found to be 100%, 602%, 0.85%, 0.18%, and 0.01%, respectively. Employing a simple extraction procedure, both test formats were applied to wheat. The detection limits for 15-AcDON in artificially contaminated wheat samples were at 50-100 ng/g; the recoveries for 15-AcDON in a concentration range from 50 to 600 ng/g were between 140.9% and 94.4%.