Catalytic hairpin-assembled and DNAzyme-driven DNA walker integrated hybridization chain reaction to construct a label-free electrochemical aptasensor for the detection of acetamiprid

As a new type of nicotine pesticide, the excessive residue of acetamiprid (ACE) in food and the environment seriously poses human health. Hence, this article developed an electrochemical aptasensor for detecting ACE based on a cascaded amplification circuit composed of catalytic hairpin-assembled and DNAzyme-driven DNA walker and hybridization chain reaction. Through the target triggered catalytic hairpin assembly (CHA), the free travelling strand of DNA walker and intact catalytic core of DNAzyme were formed, and with the assistance of Mg2+, DNAzyme was activated, driving the travelling strand to walk autonomously, repeatedly cutting hairpin DNA substrates on the electrode surface, and obtaining abundant DNA fragments. These fragments contained the initiators of hybridization chain reaction (HCR) could trigger HCR and produce the long double-strand DNA (dsDNA) for loading methylene blue (MB). Owing to the fact that Ce(Ⅲ, Ⅳ)-MOF material with electrocatalytic activity, the electrochemical reduction of MB could be effectively catalyzed, obtain significantly enhanced current signals, and ultimately achieve label-free and sensitive detection of ACE, with a detection limit of 16.38 fM (S/N = 3). Moreover, the aptasensor demonstrated excellent selectivity, stability, and reproducibility, and exhibited good reliability for detecting ACE in the vegetable soil and tomato samples, providing a new idea for the detection of pesticide residues in the fields of food and environment.