Engineering DNAzyme strategies for fluorescent detection of lead ions based on RNA cleavage-propelled signal amplification

Based on the high recognition ability and flexible programmability of GR5 DNAzyme, two fluorescent biosensors were engineered for amplified detection of Pb2+ via incorporating Ti3C2TX MXenes and embedding 2-aminopurine (2-AP), respectively. The quencher-required approach relied on the DNA affinity and fluorescence quenching ability of Ti3C2TX MXenes. Benefiting from the low background signal modulated by Ti3C2TX MXenes, the sensitive determination of Pb2+ was achieved in the linear range of 0.2-10 ng mL(-1) with the limit of detection (LOD) of 0.05 ng mL(-1). The quencher-free approach combined the fluorescent trait of 2-AP embedded in DNA structure, and the RNA cleavage-propelled digestion process of Exonuclease I (Exo I) for signal amplification, indicating the sensitive detection of Pb2+ with the LOD as low as 0.02 ng mL(-1) in the linear range of 0.1-10 ng mL(-1). Both DNAzyme assays exhibited simple procedures, favorable specificity, rapid analysis, and satisfactory application in standard reference materials (lead in drinking water) and spiked water samples. The two fluorescent biosensors established in this work would not only provide theoretic fundament for DNA adsorption of Ti3C2TX MXenes and the design of 2-AP-embedded DNAzyme assays, but also hold a great potential for on-site monitoring of lead pollution in water samples.

Automated summary

Based on the high recognition ability and flexible programmability of GR5 DNAzyme, two fluorescent biosensors were developed for amplified detection of Pb2+ using Ti3C2TX MXenes and 2-aminopurine (2-AP). The sensors demonstrated simple procedures, high sensitivity, and good specificity, making them suitable for on-site monitoring of lead pollution in water samples.