A highly sensitive electrochemical biosensor for Hg²⁺ based on entropy-driven DNA walker-based amplification

Herein, a sensitive electrochemical biosensor based on an enzyme-free and entropy-driven DNA walker is presented for the determination of Hg²⁺. This biosensor uses Hg²⁺ as a key to induce a mismatch between thymine-rich oligonucleotides to start the DNA walker, and it utilizes the entropy change of the sensing system to continuously drive the hybridization of oligonucleotides as a driving force for its walking. As the DNA walker runs, the detection signal is amplified to improve the sensitivity of the biosensor. Square wave voltammetry (SWV) of this biosensor shows a linear response of the methylene blue (MB) oxidation signal with an increase of Hg²⁺ concentration in the range of 0 to 80 nM with a detection limit of 0.136 nM, which satisfactorily meets the sensitivity requirement of the U.S. Environmental Protection Agency (EPA). The biosensor also exhibits excellent selectivity over a spectrum of interfering ions and performs well in real water samples, suggesting that it is a promising candidate for Hg²⁺ detection.