Reusable, facile, and rapid aptasensor capable of online determination of trace mercury

Herein, we reported a homemade waveguide-based evanescent wave aptasensor for the facile online monitoring of mercury pollution. The aptasensor exploited the high selectivity of hairpin structure-based thymidine-Hg²⁺-thymidine coordination chemistry (T-T mismatch) for Hg²⁺ recognition and the stably regenerable capability of DNA-functionalized waveguide surfaces. The presence of Hg²⁺ caused the T-T mismatch of Cy5.5-labeled T-rich single-stranded DNA sequences. The formed hairpin structures blocked the further hybridization of T-rich single-stranded DNA sequences with the complementary DNA strands that are modified on the waveguide surface; this phenomenon was accompanied by the decrease in the fluorescent signals excited by the evanescent wave. The limit of detection in real water samples was determined to be 0.2 μg/L, which was comparable with that of 0.4 μg/L in an ultrapure water under controlled conditions. And the linear range was observed from 1.4 µg/L to 240.7 µg/L. The negligible environmental matrix effect on the performance ensured the reliability of the proposed aptasensor. Moreover, the cross reactivity of this method toward other investigated metal ions was negligible. Through the delicate surface modification with DNA molecules covalently, the chip was reused at least 31 times with a relative standard deviation (RSD) of less than 19%. A Hg²⁺ pollution accident was successfully detected within 30 min, shedding new light in pollution monitoring, environment restoration, and emergency treatment.