Development of a sensitive, competitive, indirect ELISA for the detection of fumonisin B_1 in corn originating from Anhui province, China

摘要

Fumonisin B_1 (FB_1) 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 FB, was produced and a simple, reliable and sensitive, competitive, indirect enzyme-linked immunosorbent assay (ci-ELISA) for detection of FB_1 was developed and the experiment conditions were optimized. The coating concentration of FB,-ovalbumin (FBrOVA) was 500 ng mL~(-1), the action concentrations of anti-FB_1 mAb and goat anti-mouse IgG were 1.28 × 10~4 and 1:5000, respectively. The 50% inhibitory concentration (IC_(50)) 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 B_2 (FB_2) was 60.4, and ＜1% against deoxynivalenol (DON), aflatoxin B_1 (AFB_1), 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 FB_1 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 FB_1 in corn, furthermore, further study is needed to investigate FB_1 contamination in food and feed to prevent its harmful health effects.