Fuel strand-powered self-propelled electrochemical DNA machine for enzyme-free and distinctly amplified detection of nucleic acid with tunable performance.

In this work, a self-propelled DNA machine was proposed for enzyme-free and ultrasensitive electrochemical detection of nucleic acid, which was based on a new strategy of the fuel strand-powered target recycling and successive proximity-based inter-strand displacement for distinct signal amplification. A three-strand duplex DNA (TSD) probe was immobilized onto electrode for target DNA recognition via a fuel strand-powered strand displacement circuit, accompanied with the target recycling and the association of fuel strand onto electrode for signal amplification and readout. The associated fuel strand contained a protruding tail sequence that could be used as a target analogy to activate the neighboring TSD probe based on the proximity-based inter-strand displacement effect, inducing the self-propelled association of fuel strand onto electrode for further signal amplification. The detection performance (dynamic response range, linear interval and detection limit) of current electrochemical DNA machine could be interestingly tuned by changing the tail sequence length of fuel strand, showing the potential for different analysis requirements. The lowest detection limit of 0.1 fM could be achieved, which was lower about two orders of magnitude than that by the typical fuel strand-powered target recycling strategy. Therefore, the developed sensing system exhibits a new, simple and powerful means for amplified detection of nucleic acid and may hold great potentials in bioanalysis, disease diagnosis and biomedicine.