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Monoclonal-based enzyme-linked immunosorbent assay for the detection of zearalenone in cereals
Authors:T. Thongrussamee  N.S. Kuzmina  T. Jiratpong  S.A. Eremin  J. Intrasook
Affiliation:1. Division of Applied Life Science (Brain Korea 21 Program) , Graduate School of Gyeongsang National University, Jinju, Gyeongnam, Korea;2. Division of Applied Life Science (Brain Korea 21 Program) , Graduate School of Gyeongsang National University, Jinju, Gyeongnam, Korea;3. Mechnikov Research Institute of Vaccine and Sera of Russian Academy of Medical Sciences , Moscow, Russia;4. Chemistry Faculty, Division of Chemical Enzymology, M. V. Lomonosov Moscow State University , Moscow, Russia
Abstract:A monoclonal antibody against zearalenone (ZEA) was produced and used successfully to develop a direct competitive enzyme-linked immunosorbent assay (DC-ELISA) for the analysis of ZEA in cereals. This DC-ELISA had a limit of detection of 0.15?±?0.02 µg l?1 and an IC50 value of 1.13?±?0.16 µg l?1. Matrix interference was minimized by dilution of the sample extract before ELISA assays. Aqueous methanol (80%) gave good extraction efficiencies, and the recovery from spiked rice, barley, and corn samples averaged between 87 and 112%. Although ZEA was detected in seven (9%) of 80 rice samples and in eight (16%) of 50 barley samples, the concentration of ZEA in samples was around or below the limit of detection of DC-ELISA. Among 38 corn samples, ZEA was detected in nine (24%) samples in the range 41.0–909.8 µg kg?1. Re-analysis of the ELISA-positive corn samples by high-performance liquid chromatography (HPLC) confirmed that seven (18%) corn samples were positive. The ZEA results for corn showed very good agreement between DC-ELISA and a commercial AgraQant® zearalenone kit (r 2?=?0.98). Thus, the monoclonal antibody-based DC-ELISA could be applied to the preliminary screening of ZEA contamination when analysis of a large sample number is needed.
Keywords:zearalenone  enzyme-linked immunosorbent assay (ELISA)  immunoassay  monoclonal antibody  cereals
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