Co-contamination of mycotoxins in food and environment is a common phenomenon, which easily causes cumulative and synergistic damaging effects to human and animal health, poses great health and economy burdens to the world. A convenient technology for detection of multiple and low-concentration mycotoxins in real samples is highly desired but remains a challenge. Here, we proposed a multiplexed surface-enhanced Raman scattering (SERS)-immunochromatographic assay (ICA) that can sensitively and simultaneously detect three mycotoxins in unprocessed complex samples, using a graphene oxide-based three-dimensional (3D) Au nanofilm (called GO@Au-Au) as the film-type SERS tag. A precise sub-1 nm PEI layer was constructed into the GO@Au-Au nanostructure as built-in nanogap, which can accommodate Raman reporter molecules and create stable hotspots between the inner GO@Au film and outer assembled AuNP satellites. Grafting the 30 nm AuNPs onto the 2D GO@Au nanofilm greatly improved the SERS activity and colorimetric signal of film-type tags. Compared with traditional spherical SERS tags, the film-type GO@Au-Au can provide greater reaction interface, excellent stability and dispersibility and multiple SERS hotspots over large area for ICA using. Given these advantages, the proposed SERS-ICA can achieve simultaneous quantitative detection of fumonisin B1, aflatoxin B1, and zearalenone, with low detection limit (0.529, 0.745, and 5.90 pg mL^–1), short testing time (20 min), and high accuracy for real food/experimental samples. Our method shows great potential to fulfill practical requirements for detection of multiple mycotoxins.

食品和环境中真菌毒素的共污染是一种普遍现象，容易对人类和动物健康造成累积和协同的破坏作用，给世界带来巨大的健康和经济负担。一种用于检测真实样品中多种和低浓度霉菌毒素的便捷技术是非常需要的，但这仍然是一个挑战。在这里，我们提出了一种多重表面增强拉曼散射 (SERS)-免疫色谱法 (ICA)，它可以使用基于氧化石墨烯的三维 (3D) 金纳米膜 (称为 GO) 灵敏地同时检测未处理的复杂样品中的三种霉菌毒素@Au-Au) 作为电影类型的 SERS 标签。将精确的亚 1 nm PEI 层构建到 GO@Au-Au 纳米结构中作为内置纳米间隙，它可以容纳拉曼报告分子，并在内部 GO@Au 薄膜和外部组装的 AuNP 卫星之间产生稳定的热点。将 30 nm AuNPs 嫁接到 2D GO@Au 纳米薄膜上极大地改善了薄膜型标签的 SERS 活性和比色信号。与传统的球形 SERS 标签相比，薄膜型 GO@Au-Au 可以提供更大的反应界面、优异的稳定性和分散性以及大面积的多个 SERS 热点供 ICA 使用。鉴于这些优势，所提出的 SERS-ICA 可以实现对伏马菌素 B1、黄曲霉毒素 B1 和玉米赤霉烯酮的同时定量检测，检测限低（0.529、0.745 和 5.90 pg mL）与传统的球形 SERS 标签相比，薄膜型 GO@Au-Au 可以提供更大的反应界面、优异的稳定性和分散性以及大面积的多个 SERS 热点供 ICA 使用。鉴于这些优势，所提出的 SERS-ICA 可以实现对伏马菌素 B1、黄曲霉毒素 B1 和玉米赤霉烯酮的同时定量检测，检测限低（0.529、0.745 和 5.90 pg mL）与传统的球形 SERS 标签相比，薄膜型 GO@Au-Au 可以提供更大的反应界面、优异的稳定性和分散性以及大面积的多个 SERS 热点供 ICA 使用。鉴于这些优势，所提出的 SERS-ICA 可以实现对伏马菌素 B1、黄曲霉毒素 B1 和玉米赤霉烯酮的同时定量检测，检测限低（0.529、0.745 和 5.90 pg mL）^–1 )，测试时间短（20 分钟），对真实食品/实验样品的准确性高。我们的方法显示出满足检测多种霉菌毒素的实际要求的巨大潜力。