SPR/SERS dual-mode plasmonic biosensor via catalytic hairpin assembly-induced AuNP network

Highly sensitive and reliable detection of disease-related nucleic acids is still a big challenge in liquid biopsy because of their homologous sequences and low abundance. Herein, a novel surface plasmon resonance/surface-enhanced Raman scattering (SPR/SERS) dual-mode plasmonic biosensor based on catalytic hairpin assembly (CHA)-induced Au nanoparticle (AuNP) network was proposed for highly sensitive and reliable detection of cancer-related miRNA-652. The biosensor includes capture DNA-functionalized AuNPs (Probe 1), H1 and 4-mercaptobenzoic acid (4-MBA) co-modified AuNPs (Probe 2), and 6-carboxyl-Xrhodamine (ROX)-labeled H2 (fuel strands). The Probe 1-Probe 2 networks were formed via the target-triggered CHA reactions, which resulted in the color change of dark-field microscopy (DFM) images and enhanced SERS effect. The SPR sensing was achieved by extracting the integral optical density of dark-field color in DFM images, and the SERS sensing was realized by the ratiometric SERS signals of ROX and internal standards 4-MBA molecules. After characterizing the feasibility and optimality of the sensing strategy, the good performance of biosensors on sensitivity, specificity and uniformity was approved. The practicability of biosensors was confirmed by detecting miRNA-652 in human serum, and both the SPR and SERS assays showed good linear calibration curves and low limit of detections (LODs) of 42.5 fM and 2.91 fM, respectively, with the recovery in the range of 94.67–111.4%. These two modes show complementary advantages, and the combined SPR/SERS dual-mode can provide more options for detection and double check the results to improve the accuracy and reliability of assays, which holds a great application prospect for cancer-related nucleic acids detection in early disease stage.