DNAzyme-based magneto-controlled electronic switch for picomolar detection of lead (II) coupling with DNA-based hybridization chain reaction

摘要

A novel magneto-controlled electrochemical DNA biosensor is designed for the ultrasensitive detection of lead coupling a lead-specific DNAzyme with DNA-based hybridization chain reaction (HCR). To construct such a magnetic lead sensor, DNAzyme-based molecular beacons, selective to cleavage in the presence of Pb~(2+), are initially immobilized onto magnetic beads, which were used as the recognition elements. Upon addition of target lead, catalytic cleavage of substrate DNA segments in the double-stranded DNAzymes results in the capture of the initiator strands via the conjugated catalytic strands on magnetic beads. The captured DNA initiator strands trigger the hybridization chain reaction between two alternating hairpin DNA structures labeled with ferrocene to form a nicked double-helix on the magnetic beads. Numerous ferrocene molecules are formed on the neighboring probes, each of which produces an electrochemical signal within the applied potential. Under optimal conditions, the electrochemical signal of the magnetic lead sensor increases with the increasing lead level in the sample, and exhibits a linear response over a Pb~(2+) concentration range of 0.1-75nM with a detection limit of 37pM. Quantitative measurement of Pb~(2+) in the complex sample demonstrates the selectivity of the sensor scheme and points favorably to the application of such technologies to the analysis of environmental samples. The unique combination of a DNAzyme with hybridization chain reaction makes it possible to change the DNAzyme to select for other compounds of interest. This work represents the initial steps toward the creation of a robust field sensor for lead in groundwater or drinking water.