A three-dimensional DNA walking machine for the ultrasensitive dual-modal detection of miRNA using a fluorometer and personal glucose meter.

Three-dimensional (3D) DNA walking machines inspired by natural molecular machines have attracted significant attention due to their high walking efficiency and signal amplification capability. Herein, we report a 3D DNA walking machine for the dual-modal detection of miRNA using a fluorometer and personal glucose meter (PGM). The 3D DNA walking machine on magnetic beads (MBs) was coated with the BHQ-H1-FAM hairpin structures (DNA tracks), activated by target miRNA-21 (walking strand) and propelled by a strand displacement reaction. During these processes, the fluorescence of FAM on H1 was turned on (first signal), and the invertase on H2 was introduced into the surface of the MBs. After being separated by an external magnetic field, the invertase hydrolyzed sucrose into glucose to generate a second signal, which was quantified by the PGM. The developed 3D DNA walking machine showed high sensitivity and good specificity, and the detection limits of 98 pM and 60 pM were obtained for the fluorescence-based assay and PGM-based assay, respectively. Compared with the single-modal detection, the developed DNA walking machine can achieve a unique double signal readout and more reliable sensitive performance. In addition, the proposed 3D DNA walking machine was successfully applied to detect miRNA in real biological samples. The 3D DNA walking machine with dual-modal detection has potential applications in disease diagnostics and clinical applications and can satisfy different testing requirements both in the laboratory and field.