In-situ fabrication of 3D interior hotspots templated with a protein@Au core-shell structure for label-free and on-site SERS detection of viral diseases

Nanoscale plasmonic hotspots play a critical role in the enhancement of molecular Raman signals, enabling the sensitive and reliable trace analysis of biomedical molecules via surface-enhanced Raman spectroscopy (SERS). However, effective and label-free SERS diagnoses in practical fields remain challenging because of clinical samples' random adsorption and size mismatch with the nanoscale hotspots. Herein, we suggest a novel SERS strategy for interior hotspots templated with protein@Au core-shell nanostructures prepared via electrochemical one-pot Au deposition. The cytochrome c and lysates of SARS-CoV-2 (SLs) embedded in the interior hotspots were successfully functionalized to confine the electric fields and generate their optical fingerprint signals, respectively. Highly linear quantitative sensitivity was observed with the limit-of-detection value of 10-1 PFU/ mL. The feasibility of detecting the targets in a bodily fluidic environment was also confirmed using the proposed templates with SLs in human saliva and nasopharyngeal swabs. These interior hotspots templated with the target analytes are highly desirable for early and on-site SERS diagnoses of infectious diseases without any labeling processes.

3D interior hotspots embedded with viral lysates for rapid and label-free identification of infectious diseases

In recent decades, biomedical sensors based on surface-enhanced Raman spectroscopy (SERS), which reveals unique spectral features corresponding to individual molecular vibrational states, have attracted intensive attention. However, the lack of a system for precisely guiding biomolecules to active hotspot regions has impeded the broad application of SERS techniques. Herein, we demonstrate the irreversible active engineering of three-dimensional (3D) interior organo-hotspots via electrochemical (EC) deposition onto metal nanodimple (ECOMD) platforms with viral lysates. This approach enables organic seed-programmable Au growth and the spontaneous bottom-up formation of 3D interior organo-hotspots simultaneously. Because of the net charge effect on the participation rate of viral lysates, the number of interior organo-hotspots in the ECOMDs increases with increasingly positive polarity. The viral lysates embedded in the ECOMDs function as both a dielectric medium for field confinement and an analyte, enabling the highly specific and sensitive detection of SARS-CoV-2 lysates (SLs) at concentrations as low as 10-2 plaque forming unit/mL. The ECOMD platform was used to trace and detect the SLs in human saliva and diagnose of the delta-type SARS-CoV-2 in clinical environments;the results indicate that the proposed platform can provide point-of-care diagnoses of infectious diseases.