Development of a sensitive, competitive, indirect ELISA for the detection of fumonisin B1 in corn originating from Anhui province, China
2016
Wang, Xi C. | Bao, Ming | Li, Fu H. | Fan, Hai X. | Li, Han S. | Li, Yu | Feng, Shi B. | Wu, Jin J.
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 ₁ 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 ₁-ovalbumin (FB ₁-OVA) 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.
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