ELISA TESTS FOR DETERMINATION OF T-2 AND F2 TOXIN CONTENTS IN CEREAL SAMPLES

The authors report the setup of direct, competitive ELISA tests for de termination of Fusarium mycotoxins (T-2 and F-2) based on monoclonal a ntibodies, They analyzed the preparation of T-2- and F-2-peroxidase co njugates and optimalized the test's conditions (Fig, 2). The tests wer e validated and analyzed by important data e:g. specificity, sensitivi ty (Table I), reproducibility (Table 2) and accuracy (Table 3). The EL ISA tests were applied for quantitative determination of mycotoxins in different cereals in food and feedstuffs. Among the several extractio n solvents tried, the 89 part acetonitril/10 part 0.5% KCl/1 part 6% s ulphuric acid were chosen. The extracts can be used without further cl eanup in the ELISA test. With appropriate dilution of the sample super natants (1 :25, 1 :50), the matrix effects caused by lipid, colour, pr otein content and organic solvents (less than or equal to 10%) could b e diminished to a low level (Fig. 3). The mean recoveries from cereals infected with 100-2000 ng/g T-2 and 50-500 ng/g F-2 were 85% and 91%, respectively (Table 4 and 5). Detection limit for T-2 is 50 ng/g, tha t of F-2 is 25 ng/g. The reported reagent kits, named Toxiklon T-2 and Toxiklon Zearalenone have been commercialized. The kits are flexible and by the use of strip format microplates, individual samples in diff erent time can also be determined. In order to introduce the system in to the agricultural practice several lectures and collaborative studie s were organized for both toxins in Hungary: These kits are offered ma inly for official government laboratories for quick and quantitative a nalysis of T-2 and F-2 toxins, instead of qualitative thin-layer chrom atography in cereals.