Aptamer-Based Fluorescence Quenching Approach for Detection of Aflatoxin M-1 in Milk

文献类型： 外文期刊

第一作者： Qiao, Qinqin

作者： Qiao, Qinqin;Guo, Xiaodong;Wen, Fang;Chen, Lu;Zheng, Nan;Wang, Jiaqi;Qiao, Qinqin;Qiao, Qinqin;Cheng, Jianbo;Xue, Xiuheng;Guo, Xiaodong;Guo, Xiaodong;Wen, Fang;Chen, Lu;Zheng, Nan;Wang, Jiaqi;Xu, Qingbiao

作者机构：

关键词： aflatoxin M1; aptasensor; fluorescence; quenching; food safety

期刊名称：FRONTIERS IN CHEMISTRY （ 影响因子：3.693； 五年影响因子：4.204 ）

ISSN： 2296-2646

年卷期： 2021 年 9 卷

页码：

收录情况： SCI

摘要： Aflatoxin M-1 (AFM(1)), one of the most toxic mycotoxins, is a feed and food contaminant of global concern. In this study, we developed a fast and simple method for detection of AFM(1) based on a structure-switching signaling aptamer. This aptasensor is based on the change in fluorescence signal due to formation of an AFM(1)/aptamer complex. To generate the aptasensor, the specific aptamer was modified with FAM (carboxyfluorescein), and their complementary DNAs (cDNA) were modified with a carboxytetramethylrhodamine (TAMRA) quenching group. In the absence of AFM(1), the aptamers were hybridized with cDNA, resulting in quenching of the aptamer fluorescence due to the proximity of the aptamer's fluorophore to the quenching group on the cDNA. On the other hand, in the presence of AFM(1), a structural switch in the aptamer was induced by formation of an AFM(1)/aptamer complex. Changes in the structure of the aptamer led to the release of the cDNA, causing the generation of a fluorescence signal. Thus, AFM(1) concentrations could be quantitatively monitored based on the changes in fluorescences. Under optimized conditions, this assay exhibited a linear response to AFM(1) in the range of 1-100 ng/mL and a limit of detection of 0.5 ng/mL was calculated. This proposed aptasensor was applied to milk samples spiked with a dilution series of AFM(1), yielding satisfactory recoveries from 93.4 to 101.3%. These results demonstrated that this detection technique could be useful for high-throughput and quantitative determination of mycotoxin levels in milk and dairy products.

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