DNA-based nanomachines have aroused tremendous interest because of their potential applications in bioimaging, biocomputing, and diagnostic treatment. Herein, we constructed a novel exonuclease III-propelled and signal-amplified stochastic DNA walker that autonomously walked on a spherical particle-based 3D track through a burnt-bridge mechanism, during which nanosurface energy transfer (NSET) occurred between the fluorescent dye modified on hairpin DNA and the surface of gold nanoparticles (AuNPs). As a proof of concept, this stochastic DNA walker achieves prominent detection performance of HIV DNA in the range of 0.05-1.2 nM with a detection limit of 12.7 pM and satisfactory recovery in blood serum, showing high promise in biosensing applications with complicated media.
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