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A universal multi-wavelength fluorescence polarization immunoassay for multiplexed detection of mycotoxins in maize

机译:一种通用的多波长荧光偏振免疫分析法,用于玉米中真菌毒素的多重检测

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Multi-analyte immunoassays have attracted increasing attention due to their short assay times, low sample consumption, and reduced detection costs per assay. In this work, we describe a homologous and high-throughput multi-wavelength fluorescence polarization immunoassay (MWFPIA) for the multiplexed detection of mycotoxins. Three typical Fusarium mycotoxins, deoxynivalenol (DON), T-2 toxin and fumonisin B-1 (FB1), were labeled with different dyes. Tracers and specific monoclonal antibodies (mAbs) were employed in the MWFPIA to simultaneously detect the three mycotoxins. Under optimal conditions, the limits of detection using MWFPIA were 242.0 mu g kg(-1) for DON, 17.8 mu g kg(-1) for T-2 toxin and 331.5 mu g kg(-1) for FB1, providing sufficient sensitivity to meet the action levels of these three contaminants in maize as set by the European Union. The use of a methanol/water (2:3, v/v) mixture for sample pretreatment allowed recoveries ranging from 76.5-106.3%, with coefficients of variation less than 21.7%. The total time of analysis, including sample preparation, was less than 30 min. Twenty naturally contaminated maize samples were tested using MWFPIA and HPLC-MS/MS, with correlation coefficients (R-2) of 0.97 for DON and 0.99 for FB1. By changing the targets of interest, homologous MWFPIA, a method with high sensitivity, a simple procedure and a short analysis time, can easily be extended to other chemical contaminants. Thus, MWFPIA represents a versatile strategy for food safety analysis. (C) 2015 Elsevier B.V. All rights reserved.
机译:多分析物免疫测定法因其测定时间短,样品消耗低以及每次测定的检测成本降低而受到越来越多的关注。在这项工作中,我们描述了一种用于霉菌毒素的多重检测的同源且高通量的多波长荧光偏振免疫测定法(MWFPIA)。三种典型的镰刀菌真菌毒素,脱氧雪腐烯醇(DON),T-2毒素和伏马菌素B-1(FB1)用不同的染料标记。 MWFPIA中使用了示踪剂和特异性单克隆抗体(mAb)同时检测三种霉菌毒素。在最佳条件下,使用MWFPIA的检出限为DON为242.0μg kg(-1),T-2毒素为17.8μg kg(-1)和FB1为331.5μg kg(-1),具有足够的灵敏度满足欧盟设定的玉米中这三种污染物的作用水平。使用甲醇/水(2:3,v / v)混合物进行样品预处理可实现76.5-106.3%的回收率,变异系数小于21.7%。分析的总时间(包括样品制备)少于30分钟。使用MWFPIA和HPLC-MS / MS测试了20个自然污染的玉米样品,其中DON的相关系数(R-2)为0.97,FB1的相关系数为0.99。通过改变感兴趣的目标,同源的MWFPIA是一种灵敏度高,操作简单且分析时间短的方法,可以轻松地扩展到其他化学污染物。因此,MWFPIA代表了食品安全分析的通用策略。 (C)2015 Elsevier B.V.保留所有权利。

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