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Establishment of a Chemiluminescence Immunoassay Combined with Immunomagnetic Beads for Rapid Analysis of Ochratoxin A
Journal of AOAC INTERNATIONAL ( IF 1.7 ) Pub Date : 2021-09-30 , DOI: 10.1093/jaoacint/qsab104
Bo Zhang 1, 2 , Wenting Liu 3 , Zhenjiang Liu 1 , Xiaoling Fu 4 , Daolin Du 1
Affiliation  

Background Ochratoxin A (OTA) is a harmful mycotoxin and secondary metabolite produced by several species of Aspergillus and Penicillium. It is of vital importance to establish a sensitive method to detect OTA in various grains. Objective A novel, highly sensitive chemiluminescence immunoassay was developed to determine OTA in wheat, corn, and poultry feed. Methods The immunomagnetic beads were used as solid phase carriers and separators in the competitive chemiluminescence method. Anti-OTA monoclonal antibodies coated on the magnetic beads were applied as the capturing antibodies, in which OTA would compete with OTA-alkaline phosphatase in binding with OTA antibodies. Results The proposed method exhibited good linearity (R2 ≥ 0.999), adequate OTA recovery rate (83.60–102.50%), and good repeatability. The sensitivity of the proposed method was 2.05 pg/mL. In addition, the method was highly selective and showed no cross-reactions with other mycotoxins (e.g., AFB1, DON, ZEN). The developed method was applied in the analysis of wheat, corn, and two animal feeds successfully. The results obtained by the chemiluminescence method showed a high correlation with those obtained by the HPLC method (correlation coefficient 0.9958). Conclusion This strategy shows great potential for other toxic and harmful mycotoxin detection. Highlights A novel highly rapid and sensitive chemiluminescence immunoassay was established for quantitation of OTA.

中文翻译:

建立化学发光免疫法结合免疫磁珠快速分析赭曲霉毒素A

背景 赭曲霉毒素 A (OTA) 是一种有害的霉菌毒素和次级代谢产物,由几种曲霉属和青霉属产生。建立一种灵敏的检测各种谷物中OTA的方法至关重要。目的 开发一种新型、高灵敏度的化学发光免疫测定法,用于测定小麦、玉米和家禽饲料中的 OTA。方法采用免疫磁珠作为竞争性化学发光法中的固相载体和分离剂。将包被在磁珠上的抗OTA单克隆抗体用作捕获抗体,其中OTA将与OTA-碱性磷酸酶竞争与OTA抗体的结合。结果所提出的方法表现出良好的线性(R2≥0.999)、足够的OTA回收率(83.60-102.50%)和良好的重复性。所提出方法的灵敏度为 2。05 皮克/毫升。此外,该方法选择性高,与其他霉菌毒素(如 AFB1、DON、ZEN)无交叉反应。所开发的方法成功地应用于小麦、玉米和两种动物饲料的分析。化学发光法得到的结果与HPLC法得到的结果具有高度的相关性(相关系数为0.9958)。结论 该策略显示出其他有毒有害真菌毒素检测的巨大潜力。亮点 建立了一种用于 OTA 定量的新型高度快速和灵敏的化学发光免疫测定法。化学发光法得到的结果与HPLC法得到的结果具有高度的相关性(相关系数为0.9958)。结论 该策略显示出其他有毒有害真菌毒素检测的巨大潜力。亮点 建立了一种用于 OTA 定量的新型高度快速和灵敏的化学发光免疫测定法。化学发光法得到的结果与HPLC法得到的结果具有高度的相关性(相关系数为0.9958)。结论 该策略显示出其他有毒有害真菌毒素检测的巨大潜力。亮点 建立了一种用于 OTA 定量的新型高度快速和灵敏的化学发光免疫测定法。
更新日期:2021-09-30
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