Mycotoxin Prevention and Applied Microbiology Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Peoria, IL 61604, USA.
Toxins (Basel). 2018 Mar 10;10(3):119. doi: 10.3390/toxins10030119.
A sensitive, rapid, and reproducible imaging surface plasmon resonance (iSPR) biosensor assay was developed to detect T-2 toxin and T-2 toxin-3-glucoside (T2-G) in wheat. In this competitive assay, an amplification strategy was used after conjugating a secondary antibody (Ab₂) with gold nanoparticles. Wheat samples were extracted with a methanol/water mixture (80:20 v/v), then diluted with an equal volume of primary antibody (Ab₁) for analysis. Matrix-matched calibration curves were prepared to determine T-2 toxin and T2-G. Recovery studies were conducted at three spiking levels in blank wheat. Mean recoveries ranged from 86 to 90%, with relative standard deviations for repeatability (RSDr) of less than 6%. Limits of detection were 1.2 ng/mL of T-2 toxin and 0.9 ng/mL of T2-G, equivalent to their levels in wheat, of 48 and 36 µg/kg, respectively. The developed iSPR assay was rapid and provided enough sensitivity for the monitoring of T-2 toxin/T2-G in wheat. This is the first iSPR assay useful for detecting the "masked" T2-G in wheat.
建立了一种灵敏、快速和重现性好的表面等离子体共振(iSPR)生物传感器分析方法,用于检测小麦中的 T-2 毒素和 T-2 毒素-3-葡萄糖苷(T2-G)。在这种竞争分析中,在将第二抗体(Ab₂)与金纳米粒子偶联后,使用了一种放大策略。用甲醇/水混合物(80:20v/v)提取小麦样品,然后用相同体积的第一抗体(Ab₁)稀释进行分析。采用基质匹配校准曲线来测定 T-2 毒素和 T2-G。在空白小麦中三个添加水平进行了回收研究。平均回收率在 86%至 90%之间,重复性(RSDr)的相对标准偏差小于 6%。T-2 毒素的检测限为 1.2ng/mL,T2-G 的检测限为 0.9ng/mL,相当于其在小麦中的含量分别为 48μg/kg 和 36μg/kg。开发的 iSPR 分析方法快速且具有足够的灵敏度,可用于监测小麦中的 T-2 毒素/T2-G。这是首次用于检测小麦中“掩蔽”T2-G 的 iSPR 分析方法。