A novel visual biosensor for mercury detection based on a cleavable phosphorothioate-RNA probe and microfluidic bead trap

Mercury ion (Hg²⁺) detection plays a crucial role in safeguarding the environment and protecting human health. In this study, we developed a microfluidic biosensing platform that utilizes a phosphorothioate-RNA (PS-RNA) probe and gold-coated magnetic nanoparticles to quantify Hg²⁺ visually. The PS-RNA probe was linked to gold-coated magnetic nanoparticles via Au–S interaction and biotin–avidin interaction and then combined with polystyrene microspheres to form a self-assembled structure called a magnetic bead-AuNP-probe-polystyrene (MAPP) bead. We designed and fabricated a microfluidic chip that contained only a magnetic separator and a particle trap. The presence of Hg²⁺ cut the PS-RNA probe, leading to the separation of magnetic beads and polystyrene microspheres. The MAPP was removed by the magnetic separator, while the cut polystyrene microspheres flowed along the channel and accumulated in the particle trap, forming a long visible strip. The length of the polystyrene microsphere strip was proportional to the Hg²⁺ concentration. The microfluidic biosensing platform exhibited high sensitivity and specificity for Hg²⁺ detection, with a detection limit of 21.9 nM. By integrating the microfluidic magnetic manipulation system with a sensitive nucleic acid probe, we developed an automated and visual biosensor that is a valuable reference for the on-site visual detection of heavy metal ions.