A signal-enhancement fluorescent aptasensor based on the stable dual cross DNA nanostructure for simultaneous detection of OTA and AFB1

The simultaneous detection of multiple mycotoxins is of great significance for food safety and human health. Herein, a simple, convenient and accurate fluorescent aptasensor was designed based on the dual cross DNA nanostructure for the simultaneous detection of ochratoxin A (OTA) and aflatoxin B1 (AFB1), in which the stable dual cross DNA nanostructure provided an assay platform using the fluorescent dye-labeled aptamers as a sensing element. Owing to the higher affinity of aptamers for their target, the aptamer probes were released from the assay platform in the presence of OTA and AFB1, resulting in an enhanced fluorescence at 570 nm and 670 nm. This “signal-on” fluorescent aptasensor assay system can effectively avoid background signals and minimize false positive. Furthermore, the designed method can realize the simultaneous detection of OTA and AFB1 during the whole experiment. The limits of detection (LOD) were as low as 0.0058 ng/mL for OTA, ranging from 0.01 to 50 ng/mL and 0.046 ng/mL for AFB1, ranging from 0.05 to 100 ng/mL. The proposed fluorescent aptasensor exhibits excellent performance in practical application and provides a novel approach for the simultaneous detection of multiple mycotoxins by simply changing the aptamers. A “signal-on” fluorescent aptasensor assay system based on the stable dual cross DNA nanostructure was successfully developed for simultaneous detection of OTA and AFB1 with lower detection limits in wider linear ranges.