Inorganic mercury ion (Hg 2+ ) has been shown to coordinate to DNA duplexes that feature thymine–thymine (T–T) base pair mismatches. This observation suggests that an Hg 2+ -induced conformational change in a single-stranded DNA molecule can be used to detect aqueous Hg 2+ . Here, we have developed an analytical method using surface plasmon resonance (SPR) to develop a highly selective and sensitive detection technique for Hg 2+ that takes advantage of T–Hg 2+ –T coordination chemistry. The general concept used in this approach is that the “turn-on” reaction of a hairpin probe via coordination of Hg 2+ by the T–T base pair results in a substantial increase in the SPR response, followed by specific hybridization with a gold nanoparticle probe to amplify the sensor performance. Meanwhile, the limit of detection is 1nM, which is lower than other recently developed techniques. A linear correlation is observed between the measured SPR reflectivity and the logarithm of the Hg 2+ concentration over the concentration range of 5–5000nM. Additionally, the SPR system provides high selectivity for Hg 2+ in the presence of other divalent metal ions up to micromolar concentration levels. The proposed approach is also successfully utilized for the determination of Hg 2+ in water samples.