Methylene blue embedded duplex DNA as an efficient signal stimulator of petal-like BiVO4 for ultrasensitive photoelectrochemical bioassay

Highlights: • Methylene blue embedded in duplex DNA as hole scavengers of petal-like BiVO₄. • The developed bioassay for miRNA-21 has ultra-high sensitivity and good selectivity. • It opens up the usage of intercalated duplex DNA for efficient signal stimuli. • It offers a general strategy with wide applications in versatile PEC bioassays. Conventionally, the photoelectrochemistry relies on freely diffusive signal molecules in solution to stimulate the photocurrent output, leading to limited sensing strategies. Herein, we showcase the methylene blue (MB) embedded duplex DNA for efficient signal stimuli and its application for ultrasensitive photoelectrochemical (PEC) bioassay. Specifically, the MB embedded duplex DNA scavenged the photogenerated holes of petal-like BiVO₄ efficiently, and thus greatly augmented the anodic photocurrent output. Taking the miRNA-21 as a model target, whose biorecognition reaction was aided by the rolling circle amplification (RCA) reaction to finally produce an amplified amount of double-stranded DNA (dsDNA) with embedded MB on the photoelectrode's surface, a "label-free" and "signal-on" PEC biosensing platform was implemented with ultra-sensitivity and high selectivity. The proposed strategy could detect miRNA-21 in the concentration range of 5.0 fM to 10 nM, with the detection limit as low as 0.3 fM. This work opens up the utilization of redox substance intercalated duplex DNA for an efficient signal stimulator, which hints the prospect of other more intercalators embedded DNA for versatile biosensing purposes. Importantly, considering the large quantities of bioreactions that involve duplex DNA as reactants/products, the developed signal transduction strategy may further find wide applications in bioanalysis for targeting more analytes.