Development of a sensitive,competitive, indirect ELISA for the detection of fumonisin B1 in corn originating from Anhui province,China |
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Authors: | Xi C Wang Ming Bao Fu H Li Hai X Fan Han S Li Yu Li |
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Institution: | 1. College of Animal Science and Technology, Anhui Agricultural University, Hefei, China;2. Aquatic technology promotion station of Anhui province, Hefei, China;3. Anhui Entry-Exit Inspection and Quarantine Bureau, Hefei, China |
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Abstract: | Fumonisin B1 (FB1) is a secondary metabolite produced by Fusarium verticillioides or Fusarium proliferatum, which present in food and feed. It causes hazardous effects on human and animal health. A monoclonal antibody (mAb) against FB1 was produced and a simple, reliable and sensitive, competitive, indirect enzyme-linked immunosorbent assay (ci-ELISA) for detection of FB1 was developed and the experiment conditions were optimized. The coating concentration of FB1-ovalbumin (FB1-OVA) was 500 ng mL?1, the action concentrations of anti-FB1 mAb and goat anti-mouse IgG were 1.28 × 104 and 1:5000, respectively. The 50% inhibitory concentration (IC50) was 11 ng mL?1, with a detectable range of 1.25–250 ng mL?1, and a limit of determination (LOD) of 1.15 ng mL?1. The cross-reactivity (CR) of the antibody against fumonisin B2 (FB2) was 60.4, and <1% against deoxynivalenol (DON), aflatoxin B1 (AFB1), ochratoxin A (OTA) or zearalenone (ZEN). In spiked samples (250 ng g?1, 500 ng g?1, 1000 ng g?1), the mean recoveries ranged from 86.7 ± 5% to 102 ± 4%, and the coefficient of variation (CV) ranged from 3% to 10%. A survey of 96 corn samples from Bozhou, Fuyang, Bengbu, and Hefei, in Anhui province, China, was performed. Frequencies of FB1 contamination were 83.3%, 95.8%, 20.8% and 91.7%, and the mean concentrations of positive samples were 0.702 μg kg?1, 0.883 μg kg?1, 0.074 μg kg?1, and 0.276 μg kg?1, respectively. The results of this study suggest that the ci-ELISA developed in this study can be used to identify FB1 in corn, furthermore, further study is needed to investigate FB1 contamination in food and feed to prevent its harmful health effects. |
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Keywords: | Corn enzyme-linked immunosorbent assay (ELISA) food contamination Fumonisin B1 (FB1) monoclonal antibody (mAb) |
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