A highly sensitive and label-free microbead-based ‘turn-on’ assay was developed for the detection of Hg2+ in urine based on the Hg2+-mediated formation of intermolecular split G-quadruplex–hemin DNAzymes. In the presence of Hg2+, T–T mismatches between the two partial cDNA strands were stabilized by a T–Hg2+–T base pair, and can cause the G-rich sequences of the two oligonucleotides to associate to form a split G-quadruplex which is able to bind hemin to form the catalytically active G-quadruplex–hemin DNAzyme. This microbead-based ‘turn-on’ process allows the detection of Hg2+ in urine samples at concentrations as low as 0.5 pM. The relative standard deviation and recovery are 1.2–3.9 and 98.7–103.2%, respectively. The remarkable sensitivity for Hg2+ is mainly attributed to the enhanced mass transport ability that is inherent in homogeneous microbead-based assays. Compared with previous developments of intermolecular split G-quardruplex–hemin DNAzymes for the homogeneous detection of Hg2+ (the limit of detection was 19nM), a signal enhancement of ~1000 times is obtained when such an assay is performed on the surface of microbeads.
Comparison of DNAzyme activity for the development of an immobilized heme sensor
