SERS-based direct and sandwich assay methods for mir-21 detection

In this study, two different assay methods were developed using a surface enhanced Raman scattering (SERS) label for sensitive miR-21 detection. In the first method (direct assay), the miR-21 probe was attached to SERS-labelled, rod-shaped gold nanoparticles and hybridised with the target miR-21, which was previously immobilised onto the gold slide. In the second method (sandwich assay), the target miR-21 was captured by an miR-21 probe immobilised onto the gold slide and hybridised with a second miR-21 probe immobilised on the SERS-labeled, rod-shaped gold nanoparticles. SERS signals of developed assays were obtained via a SERS spectrum of 5,5 '-dithiobis(2-nitrobenzoic acid) (DTNB) on the rod-shaped nanoparticles. The calibration curves were plotted to measure the different concentrations of miR-21. The detection limits of the direct and sandwich assays, which last less than 40 min, were found to be 0.36 and 0.85 nM, respectively. The developed SERS-based methods offer rapid, selective, sensitive and easy detection of miR-21, especially compared to conventional PCR-based methods.