An All-in-One Aptasensor Integrating Enzyme Powered Three-Dimensional DNA Machine for Antibiotic Detection

In this work, we have developed an all-in-one aptasensor based on an enzyme-driven three-dimensional DNA walker for antibiotic detection. To overcome the drawback of time-consuming methods, high-density substrate strands were anchored on the walking interface that accelerated the signal amplification efficiency. Such an all-in-one design integrated the functionality of target recognition, signal amplification, as well as signal output into a single probe. Upon addition of kanamycin, the activated walking strand moved along the track by the stepwise cleavage of a nicking enzyme, which resulted in the enhancement of the fluorescence intensity of the solution. Under the optimized conditions, the detection process was accomplished in 40 min with a low detection limit of 1.23 pM. This aptasensor was also applied in spiked milk samples with satisfactory recoveries of 97.76% to 105.33%, demonstrating an excellent stability and accuracy. Therefore, this all-in-one aptasensor shows great potential for applications in food safety.