Exonuclease III assisted and label-free detection of mercury ion based on toehold strand displacement amplification strategy

A label-free, exonuclease III (Exo III) assisted and cost-effective Hg2+ fluorometric assay based on a strand displacement amplification strategy was developed with high sensitivity and selectivity. Thymine-thymine (T-T) mismatches between the toehold domains in hairpin DNA and assistant DNA can serve as specific recognition elements for Hg2+ binding with the help of T-Hg2+-T base pairs to initiate a toehold-mediated strand displacement reaction. Exo III provides a reduced background signal, and the method is label-free using an intercalator, Sybr green I (SG). The number of T-T mismatches, the concentration of SG and Exo III, the incubation temperature and time were all optimized. Under the optimized condition, the limit of detection for Hg2+ can be as low as 9.42 pM with a linear range from 0.01 to 1 nM. Moreover, this proposed strategy shows excellent selectivity, and was successfully applied for the detection of Hg2+ in laboratory tap water and Yanjing lake water samples in our university.