DNAzyme-Triggered Visual and Ratiometric Electrochemiluminescence Dual-Readout Assay for Pb(II) Based on an Assembled...

Currently, portable, low-cost, and easy to operate on-chip analytical units are urgently demanded to meet the requirement for point-of-care testing in resource-limited regions. Herein, a dual-mode lab-on-paper platform is presented, which integrates distance-based visualized readout with ratiometric electrochemiluminescence (ECL) assay in one device. The distance-based measurement is based on a brown visualized strip generated from the oxidation reaction of 3,3'-diaminobenzidine in the presence of H₂O₂ initiated by horseradish peroxidase (HRP). Notably, visualized semiquantitative results are displayed as the length of a brown bar chart directly on the device-without the need for any data processing or plotting steps, thus avoiding the error caused by the naked eye for distinguishing the color depth. On the contrary, a ratiometric ECL technique was employed for accurate analysis based on the specific biorecognition between Pb²⁺-dependent DNAzymes and targets. Concretely, upon addition of Pb²⁺ into the fabricated device, cleaved oligonucleotide fragments connected with HRP functionalized Au nanocubes could permeate through the cellulose on account of their size that is smaller than paper pores, quench the ECL signal of the CdS quantum dots because of resonance energy transfer, and synchronously boost the ECL intensity generated from luminol by catalyzing H₂O₂. As a consequence, satisfied prediction and accurate monitoring performance was obtained in the range 0.1-2000 nM and 0.01-2000 nM by measuring the length of colored product and the ratio of ECL intensity, respectively. The beneficial advantages of low cost, high efficiency, and the capacity to perform dual-mode assay qualify this innovative device for use with diverse applications.