A rapid, sensitive and multiplexed imaging surface plasmon resonance (iSPR) biosensor assay was developed and validated for three Fusarium toxins, deoxynivalenol (DON), zearalenone (ZEA) and T-2 toxin. The iSPR assay was based on a competitive inhibition format with secondary antibodies (Ab₂) conjugated to gold nanoparticles (AuNPs) used as a signal amplification tag. Signal was amplified nearly 25-fold for DON, 90-fold for ZEA and 12-fold for T-2 toxin assay using Ab₂-AuNPs. Analyses, including steps to regenerate the sensor, took 17.5 min. The antigen coated sensor chip was used for more than 46 cycles without affecting signal intensity (< 12%). Matrix matched calibration curves were used to determine Fusarium toxins in wheat. The mean recoveries ranged from 87% to 103% with relative standard deviations of repeatability of less than 5%. The limits of detection were 15 µg/kg for DON, 24 µg/kg for ZEA and 12 µg/kg for T-2 toxin. This provided sufficient sensitivity to monitor contamination of these mycotoxins in wheat in accordance with European Commission (EC) limits. Cut off levels for all three Fusarium toxins were validated using blank wheat and wheat spiked either at the EC regulated levels (100 µg/kg for ZEA and T-2 toxin) or at one third of the EC level (for DON: 400 µg/kg). The assay was successfully applied and further validated with naturally contaminated wheat samples. This is the first reported AuNP enhanced iSPR assay to detect and classify three agriculturally important Fusarium toxins in wheat.

快速，灵敏和多重成像表面等离振子共振（iSPR）生物传感器测定方法已经开发出来，并针对三种镰刀菌毒素，脱氧雪腐酚（DON），玉米赤霉烯酮（ZEA）和T-2毒素进行了验证。iSPR分析基于竞争性抑制形式，其中偶联至金纳米颗粒（AuNPs）的_二抗（Ab ₂）被用作信号放大标签。使用Ab ₂ -AuNPs ，DON的信号放大了近25倍，ZEA的信号放大了90倍，T-2毒素检测的信号放大了12倍。分析（包括重新生成传感器的步骤）花费了17.5分钟。使用抗原包被的传感器芯片进行了46个以上的循环，而不会影响信号强度（<12％）。使用基质匹配的校准曲线确定镰刀菌小麦中的毒素。平均回收率从87％到103％不等，重复性的相对标准偏差小于5％。DON的检出限为15 µg / kg，ZEA的检出限为24 µg / kg，T-2毒素的检出限为12 µg / kg。根据欧洲委员会（EC）的限制，这提供了足够的敏感性来监测小麦中这些真菌毒素的污染。使用空白小麦和掺加了EC规定水平（ZEA和T-2毒素为100 µg / kg）或EC水平的三分之一（DON：DON为400 µg /）的小麦验证了所有三种镰刀菌毒素的截断水平。公斤）。该测定法已成功应用，并用天然污染的小麦样品进行了进一步验证。这是第一个报道的AuNP增强iSPR测定法，用于检测和分类三个农业上重要的镰刀菌 小麦中的毒素。