Sensitive label-free oligonucleotide-based microfluidic detection of mercury (II) ion by using exonuclease I

Abstract Mercury is a highly toxic metal that can cause significant harm to humans and aquatic ecosystems. This paper describes a novel approach for mercury (Hg 2+ ) ion detection by using label-free oligonucleotide probes and Escherichia coli exonuclease I (Exo I) in a microfluidic electrophoretic separated platform. Two single-stranded DNAs (ssDNA) TT-21 and TT-44 with 7 Thymine–Thymine mispairs are employed to capture mercury ions. Due to the coordination structure of T–Hg 2+ –T, these ssDNAs are folded into hairpin-like double-stranded DNAs (dsDNA) which are more difficult to be digested by Exo I, as confirmed by polyacrylamide gel electrophoresis (PAGE) analysis. A series of microfluidic capillary electrophoretic separation studies are carried out to investigate the effect of Exo I and mercury ion concentrations on the detected fluorescence intensity. This method has demonstrated a high sensitivity of mercury ion detection with the limit of detection around 15 nM or 3 ppb. An excellent selectivity of the probe for mercury ions over five interference ions Fe 3+ , Cd 2+ , Pb 2+ , Cu 2+ and Ca 2+ is also revealed. This method could potentially be used for mercury ion detection with high sensitivity and reliability.