SERS-based sandwich bioassay protocol of miRNA-21 using Au@Ag core-shell nanoparticles and a Ag/TiO₂ nanowires substrate

Based on surface-enhanced Raman scattering (SERS) technology, Au@Ag@4MBA@5′-NH₂-ssDNA probes and a Ag/TiO₂@3′-NH₂-ssDNA substrate were prepared and constructed into a sandwich structure to develop a high sensitivity bioassay of miRNA-21. The Au@Ag@4MBA@5′-NH₂-ssDNA probes were prepared by immobilizing 5′-NH₂-ssDNA onto the surfaces of 4MBA-labelled Au@Ag core-shell nanoparticles, and the Ag/TiO₂@3′-NH₂-ssDNA substrate was prepared by immobilizing 3′-NH₂-ssDNA on the surface of Ag/TiO₂ nanowires SERS-active substrates. The experimental results showed that the SERS-based sandwich bioassay of miRNA-21 presented a low limit of detection of 0.75 fM and a broad dynamic range from 1.0 fM to 1.0 nM. Also, the test data for the SERS-based sandwich bioassay were not only consistent with that of the real-time fluorescence quantitative polynucleotide chain reaction (RT-qPCR) method but also displayed higher detection sensitivity. It was shown that the SERS-based sandwich bioassay of miRNA-21 has importance for use in potential applications involving diagnosing clinical cancer patients.