Ultrasensitive colorimetric aptasensor for Hg²⁺ detection using Exo-III assisted target recycling amplification and...

Facile and ultrasensitive detection of Hg²⁺ in water environment remains challenging. Exonuclease III (Exo-III)-assisted target recycling is one of the most popular amplification strategies. Although the magnesium (II) ions are widely acting as cofactors of Exo-III, we recognized that Mg²⁺ cofactors would strongly disturb the charge distribution on citrate-stablized gold nanoparticles (in the general sense, unmodified AuNPs) surface, thus generate false positive colorimetric signals. To address this issue, we first put forward the view that the cobalt (II) ions can function as the Exo-III cofactor and successfully construct a novel label-free colorimetric aptasensor for facile and ultrasensitive detection of Hg²⁺ using Hg²⁺-triggered Exo-III-assisted signal amplification and unmodified AuNPs as indicators. A hairpin-looped DNA probe was rationally designed with thymine-rich recognition termini and specifically recognized trace Hg²⁺ by a stable T-Hg²⁺-T structure. A blue-to-red color change of AuNPs with the addition of Hg²⁺ provided the quantitative detection of Hg²⁺ with a limit of detection of 0.2 nM and a linear working range from 0.5 nM to 5.0 nM. The whole testing time for one assay was approximately 40 min. Real water samples, even containing Hg²⁺ at 1 nM, could be determined by the aptasensor with recovery rates from 97% to 103%.