Journal
ANALYTICAL METHODS
Volume -, Issue -, Pages -Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/d3ay01661g
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This paper presents a comprehensive study on the design and implementation of a SERS biosensor for detecting the specific miRNA biomarker miRNA-183 for COPD. By utilizing a labeled molecular beacon and a specially designed MB, the biosensor achieves high sensitivity and specificity in detecting target molecules at femtomolar concentrations, and the sensor's regenerability is also demonstrated.
Advantages of biosensors based on surface enhanced Raman scattering (SERS) rely on improved sensitivity and specificity, and suited reproducibility in detecting a target molecule that is localized in close proximity to a SERS-active surface. Herein, a comprehensive study on the realization of a SERS biosensor designed for detecting miRNA-183, a miRNA biomarker that is specific for chronic obstructive pulmonary disease (COPD), is presented. The used strategy exploits a signal-off mechanism by means of a labelled molecular beacon (MB) as the oligonucleotide biorecognition element immobilized on a 2D SERS substrate, based on spot-on silver nanowires (AgNWs) and a multi-well low volume cell. The MB was properly designed by following a dedicated protocol to recognize the chosen miRNA. A limit of detection down to femtomolar concentration (3 x 10-16 M) was achieved and the specificity of the biosensor was proved. Furthermore, the possibility to regenerate the sensing system through a simple procedure is shown: with regeneration by using HCl 1 mM, two detection cycles were performed with a good recovery of the initial MB signal (83%) and a reproducible signal after hybridization. To overcome the issues connected to the need of multiple steps in amplification methods for SERS biosensors, we propose a strategy for microRNA detection based on a signal-off mechanism of a labelled molecular beacon immobilized on the SERS substrate.
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