State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing, 100193, China.
College of Veterinary Medicine, China Agricultural University, Beijing, China.
J Hazard Mater. 2020 Jul 5;393:122348. doi: 10.1016/j.jhazmat.2020.122348. Epub 2020 Feb 19.
A multiplex surface-enhanced Raman scattering (SERS)-based lateral flow immunosensor was developed to determine six major mycotoxins in maize. Two characteristic Raman reporter molecules-5,5-dithiobis-2-nitrobenzoic acid (DTNB) and 4-mercaptobenzoic acid (MBA)-were used to label the synthesized Au@Ag core-shell nanoparticles for the preparation of SERS nanoprobes as detection reagents. Six corresponding hapten-protein conjugates were prepared and dispensed on three test lines of nitrocellulose membrane with two conjugates on each line as capture antigens. This design facilitates the simultaneous detection of the six mycotoxins in a single test. After optimizing the experimental parameters of immunosensor, the limits of detection were as low as 0.96 pg/mL for aflatoxin B, 6.2 pg/mL for zearalenone, 0.26 ng/mL for fumonisin B, 0.11 ng/mL for deoxynivalenol, 15.7 pg/mL for ochratoxin A, and 8.6 pg/mL for T-2 toxin, respectively. The spiking experiment showed high accuracy with recovery of 78.9-106.2 % and satisfactory assay precision with the coefficient of variations below 16 %. Moreover, this assay can be completed in less than 20 min, and its detection results were consistent with that of liquid chromatography-mass spectrometry. Therefore, the developed SERS-based lateral flow immunosensor is a promising approach for mycotoxin detection in the field.
建立了一种多重表面增强拉曼散射（SERS）侧向流免疫传感器，用于测定玉米中的六种主要真菌毒素。两种特征拉曼报告分子-5,5-二硫代双（2-硝基苯甲酸）（DTNB）和 4-巯基苯甲酸（MBA）-被用于标记合成的 Au@Ag 核壳纳米粒子，以制备 SERS 纳米探针作为检测试剂。制备了六种相应的半抗原-蛋白缀合物，并分配在硝酸纤维素膜的三个测试线上，每个线上有两个缀合物作为捕获抗原。这种设计便于在单次测试中同时检测六种真菌毒素。在优化免疫传感器的实验参数后，检测限低至黄曲霉毒素 B 为 0.96 pg/mL，玉米赤霉烯酮为 6.2 pg/mL，伏马菌素 B 为 0.26 ng/mL，脱氧雪腐镰刀菌烯醇为 0.11 ng/mL，赭曲霉毒素 A 为 15.7 pg/mL，T-2 毒素为 8.6 pg/mL。加标实验显示回收率为 78.9-106.2%，具有较高的准确性，变异系数低于 16%，具有良好的测定精密度。此外，该测定方法可在 20 分钟内完成，其检测结果与液相色谱-质谱法一致。因此，开发的基于 SERS 的侧向流免疫传感器是一种很有前途的现场真菌毒素检测方法。
