Triple-Helix Molecular Switch Triggered Cleavage Effect of DNAzyme for Ultrasensitive Electrochemical Detection of Chloramphenicol

The abuse of chloramphenicol (CAP) in animal-derived products leads to serious food safety problems, so the sensitive and accurate determination of CAP residues has great noteworthiness for public health. Herein, we present a novel electrochemical aptasensor that incorporates a poly-(diallyldimethylammonium chloride) functionalized graphene/Ag@Au nanosheets (PDDA-Gr/Ag@Au NSs) composite modified electrode and a DNAzyme signal amplification effect triggered by a triple-helix molecular switch (THMS) for detecting CAP. The PDDA-Gr/Ag@Au NSs composite has the advantages of high surface area, great conductivity, and dispersibility and has successfully improved the electrochemical performance of the electrode. Specific interaction with CAP will cause the signal transduction probe (STP) to be released from the THMS. After that, the DNAzyme will be activated with the help of Pb2+ and remove the immobilized signal probe on the electrode surface. The signal change was recorded by square wave voltammetry (SWV) and led to an accurate quantification of CAP. With all these features, the proposed sensing strategy yielded a satisfactory analytical performance with linearity between 1 pM and 1 mu M and a limit of detection of 18.6 fM. Furthermore, the aptasensor shows excellent specificity for CAP in the presence of other antibiotics and resists interference with other common metal ions. Importantly, the performance is not diminished when the constructed aptasensor is applied to measuring CAP in milk powder. This THMS-based method is easy to design, and alteration to different targets can be achieved by simply replacing the aptamer sequence in the THMS. Therefore, this method shows significant prospects as a flexible platform for accurate monitoring of antibiotic residues in foodstuffs.

Automated summary

In this study, a novel electrochemical aptasensor was developed for sensitive and accurate determination of chloramphenicol (CAP) residues in animal-derived products. The aptasensor incorporated a poly-(diallyldimethylammonium chloride) functionalized graphene/Ag@Au nanosheets composite modified electrode and a DNAzyme signal amplification effect triggered by a triple-helix molecular switch (THMS). The aptasensor showed high sensitivity and specificity for CAP, and was not affected by interference from other antibiotics and common metal ions. The method also performed well when applied to measuring CAP in milk powder.