Fluorometric determination of mercury(II) by using thymine-thymine mismatches as recognition elements, toehold binding, and enzyme-assisted signal amplification.

A highly sensitive fluorometric method is described for the determination of mercury(II) ions. It is based on (a) the use of a DNA probe containing thymine-thymine mismatches that are employed as Hg(II) recognition elements, (b) subsequent toehold binding, and (c) endocuclease-assisted signal amplification. Target recycling is triggered by exonuclease III. This produces a large amount of ssDNA (defined as primer). Then, the generated primer-initiated strand displacement reaction with the help of polymerase and nicking endonuclease releases the free fluorophore-labelled probe. Under excitation at 532 nm, the fluorescent probe displays emission with a peak at 582 nm. The sensitivity of this method is improved by introduction of nicking endonuclease. The working range of the assay extends from 20 pM to 10 nM, and the detection limit is as low as 6 pM of Hg(II). Graphical abstract Schematic presentation of the fluorometric method for determination of mercury(II). By using a special structure of thymine-thymine mismatches, target-induced toehold binding and enzyme-assisted signal amplification strategy were employed. This method is selective and good performance in real sample application.