A bivalent binding aptamer-cDNA on MoS₂ nanosheets based fluorescent aptasensor for detection of aflatoxin M₁

To ensure the safety of dairy products, especially milk, and consequently protect human health, accurate and simple analytical techniques are highly necessary to determine the low concentration of aflatoxin M₁ (AFM₁) as an important carcinogen. Herein, a novel, accurate and simple fluorescent aptasensor was designed for selective detection of AFM₁ based on bivalent binding aptamer-cDNA (BBA-cDNA) structure. Moreover, MoS₂ nanosheets (MoS₂ NSs) were used as the fluorescent quencher and FAM-labeled complementary strand of aptamer (FAM-CS) was applied as a fluorescent probe. In this study, we achieved a new result. Unlike previous studies, in this work, the BBA-cDNA structure was not disassembled in the presence of the target. Therefore, as the AFM1 concentration increased, more targets were attached to the BBA-cDNA structure and as a result, the BBA-cDNA structure/AFM1 could not be placed on the surface of MoS2 NSs, leading to the more fluorescent intensity detection. Under optimized conditions, the developed fluorescent analytical method revealed great selectivity toward AFM₁ with a limit of detection (LOD) of 0.5 nM and a linear range from 0.7 to 10 nM. This fabricated aptasensor indicated excellent analytical performance for AFM₁ detection in milk samples with LOD of 0.1 nM. Overall, the proposed approach could provide an effective basis for small molecule analysis to guarantee food and human safety using appropriate aptamer sequences.