Fluorescent detection of Cu (II) ions based on DNAzymatic cascaded cyclic amplification

A fluorescent biosensor based on the cascaded cyclic amplification-lighted copper nanoparticles has been developed, optimized, and validated. In the double-modular cascaded cyclic amplification, a DNAzymatic cyclic amplification unit transforms metal ion signal to specific DNA sequences, and a linear/exponential integrated amplification unit converts as-prepared DNA codes to identical thymine (T)-rich DNA templates. T-rich scaffolds can induce the generation of red fluorescent copper nanoparticles, with fluorescence emission at 625 nm upon the excitation at 340 nm, as signal vehicles for quantitative detection of metal ions. Copper ions, selected as the model target, could be detected in a wide linear range from 10 to 10⁴ nM depending on the increased fluorescent intensity, and the detection limit is 5.6 ± 0.52 nM (n = 3) within 40 min, which is 4 orders of magnitude lower than the limits set in drinking water. In the detection of Cu²⁺ in real tap and lake water, the results between inductively coupled plasma mass spectrometry (ICP-MS) and our proposed biosensor were consistent, illustrating the practicability of the fabricated method. In summary, the established fluorescent biosensor compensates the deficiency of immunoassays failing to analyze metal ions, broadens ranges of biomarkers responding to cleaved DNAzymes, provides an open platform sensing different metal ions, and meets the increasing need for the ultrasensitive detection in the field of food safety, environmental monitoring, and medical diagnosis.