A surface plasmon resonance (SPR)-enhanced optical signal using a nanoslit array and acridine orange (AO) dye system at a flexible poly(dimethylsiloxane) (PDMS) substrate was achieved in this work and demonstrated a simple sensing scheme to directly detect SARS-CoV-2 nucleic acid via DNA hybridization. A simple nanoimprinting pattern transfer technique was introduced to form uniform reproducible nanoslit arrays where the dimensions of the slit array were controlled by the thickness of the gold film. The plasmon–exciton coupling effect on the optical enhancement of different dye molecules, i.e., AO, propidium iodide (PI), or dihydroethidium (DHE) attached to the nanoslit surfaces, was examined thoroughly by measuring the surface reflection and fluorescence imaging. The results indicate that the best overlap of the plasmon resonance wavelength to the excitation spectrum of AO presented the largest optical enhancement (∼57×) compared to the signal at flat gold surfaces. Based on this finding, a sensitive assay for detecting DNA hybridization was generated using the interaction of the selected SARS-CoV-2 ssDNA and dsDNA with AO to trigger the metachromatic behavior of the dye at the nanoarray surfaces. We found strong optical signal amplification on the formation of acridine-ssDNA complexes and a quenched signal upon hybridization to the complementary target DNA (ct-DNA) along with a blue shift in the fluorescence of AO-dsDNAs. A quantitative evaluation of the ct-DNA concentration in a range of 100–0.08 nM using both the reflection and emission imaging signals demonstrated two linear regimes with a lowest detection limit of 0.21 nM. The sensing method showed high sensitivity and distinguished signals from 1-, 2-, and 3-base mismatched DNA targets, as well as high stability and reusability. This approach toward enhancing optical signal for DNA sensing offers promise in a general, rapid, and direct vision detection method for nucleic acid analytes.

Here we present a plasmonic nanoledge device with high sensitivity and selectivity used to detect protein biomarkers simply by functionalizing the device, which specifically binds to particular biomolecule or biomarkers. We employ this plasmonic nanoledge device for the detection of anti-insulin antibodies of type 1 diabetes (T1D) in buffer and human serum at the range of pg ml −1 to 100 ng ml −1 . The signal transduction is based on the extraordinary optical transmission (EOT) through the nanoledge array and the optical spectral changes with the biological binding reaction between the surface functionalized insulin with anti-insulin antibody. Control experiments indicate little interferences from the human serum background and addition of other proteins such as bovine serum albumin (BSA) and epidermal growth factor (EGF) at 20 ng ml −1 . The high sensitivity, specificity and easy adaptability of the plasmonic device offer new opportunities in biosensing and diagnostic applications for T1D.

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