Henan Engineering Technology Research Center of Food Processing and Circulation Safety Control, College of Food Science and Technology, Henan Agricultural University, Zhengzhou, 450002, Henan, China; Henan Technology Innovation Center of Meat Processing and Research, College of Food Science and Technology, Henan Agricultural University, Zhengzhou, 450002, Henan, China.
Henan Engineering Technology Research Center of Food Processing and Circulation Safety Control, College of Food Science and Technology, Henan Agricultural University, Zhengzhou, 450002, Henan, China; Henan Technology Innovation Center of Meat Processing and Research, College of Food Science and Technology, Henan Agricultural University, Zhengzhou, 450002, Henan, China; International Education College, Henan Agricultural University, Zhengzhou, 450002, Henan, China.
Talanta. 2023 Dec 1;265:124908. doi: 10.1016/j.talanta.2023.124908. Epub 2023 Jul 6.
Realizing the simultaneous speedy detection of multiple mycotoxins in contaminated food and feed is of great practical importance in the domain of food manufacturing and security. Herein, a fluorescent aptamer sensor based on self-assembled DNA double-crossover was developed and used for effective simultaneous quantitative detection of aflatoxins M and B by fluorescence resonance energy transfer (FRET). Fluorescent dye-modified aflatoxin M and B aptamers are selected as recognition elements and signal probes, and DNA double crosses are consistently locked by the aflatoxin aptamers, which results in a "turn-off" of the fluorescent signal. In the presence of AFM and AFB, the aptamer sequences are more inclined to form Apt-AFM and Apt-AFB complexes, and the fluorescent probes are released from the DNA double-crossing platform, leading to an enhanced fluorescent signal (Cy3: 568 nm; Cy5: 660 nm). Under the optimal conditions, the signal response of the constructed fluorescent aptamer sensor showed good linearity with the logarithm of AFM and AFB concentrations, with detection limits of 6.24 pg/mL and 9.0 pg/mL, and a wide linear range of 0.01-200 ng/mL and 0.01-150 ng/mL, respectively. In addition, the effect of potential interfering substances in real samples was analyzed, and the aptasensor presented a good interference immunity. Moreover, by modifying and designing aptamer probes, the sensor can be applied to high-throughput simultaneous screening of other analytes, providing a new approach for the development of fluorescent aptamer sensors.
实现对污染食品和饲料中多种真菌毒素的同时快速检测，在食品制造和安全领域具有重要的实际意义。在此，我们开发了一种基于自组装 DNA 双交叉的荧光适体传感器，并将其用于通过荧光共振能量转移（FRET）对黄曲霉毒素 M 和 B 进行有效的同时定量检测。荧光染料修饰的黄曲霉毒素 M 和 B 适体被选为识别元件和信号探针，并且 DNA 双交叉始终被黄曲霉毒素适体锁定，导致荧光信号“关闭”。在存在 AFM 和 AFB 的情况下，适体序列更倾向于形成 Apt-AFM 和 Apt-AFB 复合物，并且荧光探针从 DNA 双交叉平台释放，导致荧光信号增强（Cy3：568nm；Cy5：660nm）。在最佳条件下，所构建的荧光适体传感器的信号响应与 AFM 和 AFB 浓度的对数呈良好的线性关系，检测限分别为 6.24pg/mL 和 9.0pg/mL，线性范围分别为 0.01-200ng/mL 和 0.01-150ng/mL。此外，分析了实际样品中潜在干扰物质的影响，适体传感器表现出良好的抗干扰能力。此外，通过修饰和设计适体探针，该传感器可应用于其他分析物的高通量同时筛选，为荧光适体传感器的发展提供了一种新方法。
