Ultrasensitive colorimetric strategy for Hg²⁺ detection based on T-Hg²⁺-T configuration and target recycling amplification

A novelty aptasensor for ultrasensitive detection of Hg²⁺ is developed, exploiting the combination of plasmonic properties of gold nanoparticles (AuNPs) and exonuclease III (Exo III)-assisted target recycling for signal amplification. In the presence of Hg²⁺, a DNA duplex can be formed due to the strong coordination of Hg²⁺ and T bases of single-stranded DNA (ssDNA) probe. Exo III digests the DNA duplex from the 3′ to 5′ direction, resulting in the releasing of Hg²⁺. Then, the released Hg²⁺ binds with another ssDNA probe through T-Hg²⁺-T coordination. After Exo III-assisted Hg²⁺ cycles, numerous ssDNA probes are exhausted, which promotes poly(diallyldimethylammonium chloride) (PDDA)-induced AuNP aggregation, leading to an obvious color change and aggregation-induced plasmon red shift of AuNPs (from 520 to 610 nm). Therefore, this biosensor is ultrasensitive, which is applicable to the detection of trace level of Hg²⁺ with a linear range from 5 pM to 0.6 nM and an ultralow detection limit of 0.2 pM. Furthermore, it enables visual detection of Hg²⁺ as low as 50 pM by the naked eye. More importantly, the assay can be applied to the reliable determination of spiked Hg²⁺ in sea water samples with good recovery.