title: A ratiometric electrochemical biosensor for sensitive detection of Hg2+ based on thymine-Hg2+-thymine structure.
abstract: In this paper, a simple, selective and reusable electrochemical biosensor for the sensitive detection of mercury ions (Hg(2+)) has been developed based on thymine (T)-rich stem-loop (hairpin) DNA probe and a dual-signaling electrochemical ratiometric strategy. The assay strategy includes both "signal-on" and "signal-off" elements. The thiolated methylene blue (MB)-modified T-rich hairpin DNA capture probe (MB-P) firstly self-assembled on the gold electrode surface via Au-S bond. In the presence of Hg(2+), the ferrocene (Fc)-labeled T-rich DNA probe (Fc-P) hybridized with MB-P via the Hg(2+)-mediated coordination of T-Hg(2+)-T base pairs. As a result, the hairpin MB-P was opened, the MB tags were away from the gold electrode surface and the Fc tags closed to the gold electrode surface. These conformation changes led to the decrease of the oxidation peak current of MB (IMB), accompanied with the increase of that of Fc (IFc). The logarithmic value of IFc/IMB is linear with the logarithm of Hg(2+) concentration in the range from 0.5 nM to 5000 nM, and the detection limit of 0.08 nM is much lower than 10nM (the US Environmental Protection Agency (EPA) limit of Hg(2+) in drinking water). What is more, the developed DNA-based electrochemical biosensor could be regenerated by adding cysteine and Mg(2+). This strategy provides a simple and rapid approach for the detection of Hg(2+), and has promising application in the detection of Hg(2+) in real environmental samples.
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
authors.forename: Erhu
authors.initials: E
authors.lastname: Xiong
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
authors.forename: Liang
authors.initials: L
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
authors.forename: Jiawan
authors.initials: J
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
authors.forename: Peng
authors.initials: P
authors.lastname: Yu
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China. Electronic address: mickyxie@hnu.edu.cn.
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China. Electronic address: chenjinhua@hnu.edu.cn.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Chen
mesh_terms: D015374:Biosensing Techniques; D015342:DNA Probes / Q000737:chemistry; D060766:Drinking Water / Q000737:chemistry; D055664:Electrochemical Techniques; D004566:Electrodes; D006046:Gold / Q000737:chemistry; D006801:Humans; D007477:Ions / Q000737:chemistry; D008628:Mercury / Q000032:analysis* / Q000097:blood / Q000737:chemistry; D008751:Methylene Blue / Q000737:chemistry; D010084:Oxidation-Reduction; D013941:Thymine / Q000737:chemistry*
chemical_list: D015342:DNA Probes; D060766:Drinking Water; D007477:Ions; D006046:Gold; D008628:Mercury; D013941:Thymine; D008751:Methylene Blue
keywords: Electrochemistry; Ferrocene; Mercury ion; Methylene blue; Ratiometric biosensor