A dual-model "on-super off" photoelectrochemical/ratiometric electrochemical biosensor for ultrasensitive and accurate detection of microRNA-224.

A dual-model "on-super off" photoelectrochemical (PEC)/ratiometric electrochemical (EC) biosensor based on signal enhancing and quenching combining three-dimensional (3D) DNA walker strategy was designed for the ultrasensitive and accurate detection of microRNA-224 (miRNA-224). The "signal on" PEC state was achieved by methylene blue labeled hairpin DNA (MB-DNA) for sensitizing CdS QDs. Then numerous transformational ferrocene labeled DNAs (Fc-DNAs) converted by target-induced 3D DNA walker amplification with the help of Ag nanocubes (NCs) label DNA (Ag-DNA) were introduced to open hairpin MB-DNA. Such configuration change would relocate the sensitizer MB and the quencher Fc, whereas energy transfer placed between Ag NCs and CdS QDs, thereby significantly quenching the PEC signal to obtain "super off" state. Meanwhile, these changes resulted in a decreased oxidation peak current of MB (I) and an increased that of Fc (I). MiRNA-224 was also detected on basis of the dual-signaling EC ratiometric method for complementary PEC detection. Benefiting from different mechanisms and relatively independent signal transduction, this approach not only avoided interference from difficult assembly but also outstandingly increased sensitivity by distance-controllable signal enhancing and quenching strategies. As a result, the detection ranges of 0.1-1000 fM with a low detection limit of 0.019 fM for PEC, and 0.52 to 500 fM with a low detection limit of 0.061 fM for EC, were obtained for miRNA-224, which opens a new avenue for designing numerous elegant biosensors with potential utility in bioanalysis and early disease diagnosis.