A novel electrochemical Hg(2+) biosensor was developed on the basis of a metal-organic hybrid microarray, in which the nicking endonuclease (NE) assisted target-triggered strand release strategy was realized via the DNA cyclic amplification technique. The metal-organic hybrid microarray was fabricated using the SAM of 1, 4-benzenendithiol as soft template, and the density of the microarray could be adjusted by controlling the surface coverage of 1,4-benzenendithiol molecules. In the presence of Hg(2+), capture DNA (cDNA) with an indicator at one end could hybridize with the reporter DNA (rDNA) through the stable T-Hg(2+)-T linkage, forming the nicking recognition site. After the nicking reaction, the electrochemical indicator dissociated from the electrode surface. The released rDNA and Hg(2+) could be reused in the sensing system and initiate the next cycle, and more electroactive indicator dissociated from the electrode surface, resulting in a significant signal decrease. The constructed DNA biosensor could detect Hg(2+) in a wide linear range from 15 pM to 500 nM, with an ultrasensitive detection limit of 5 pM (S/N=3). Furthermore, the biosensor exhibited excellent stability, good reproducibility and high selectivity towards other divalent ions. The proposed sensing system also showed a promising potential for the application in real aquatic product sample analysis.
Keywords: DNA cyclic amplification; Electrochemical DNA biosensor; Hg(2+) detection; Metal-organic hybrid microarray; Nicking endonuclease.
© 2013 Published by Elsevier B.V.