Signal-amplified surface-enhanced Raman scattering using core/shell satellite nanoparticles for norovirus detection

The development of an innovative approach is explored to amplify the signal of a surface-enhanced Raman scattering (SERS)-based detection system using a novel nanotag: Au@Ag NPs covered by satellite AuNPs and conjugated by 4-mercaptbenzoic acid (4-MBA) as a Raman tag (Au@Ag-MBA-AuNPs). The Au@Ag-MBA-AuNPs nanotags showed strong SERS activities with an enhancement factor in the 10⁸ order of magnitude. This indicates the formation of many hot spots due to the combination of core-shell nanoparticles and satellite AuNPs on the surface of Au@Ag-MBA NPs. The newly fabricated nanotags were employed in a small-sized Palmtop Raman spectrometer. A concentration-dependent increase in SERS intensity was observed in the norovirus-like particle (NoV-LP) concentration range 10 fg/mL to 100 pg/mL with a detection limit of 0.76 fg/mL. Even in the severe interfering matrices, this detection method's coefficient of variation was less than 10%. This detection system was approximately 10⁷ times more sensitive than commercially available ELISA kits. Norovirus in clinical samples was detected over a wide concentration range of 1.0 × 10¹ - 1.0 × 10⁶ RNA copy number/mL with a detection limit of 7.8 RNA copy number/mL, indicating sensitivity comparable to real-time PCR. These results suggest that this detection system is stable in a complex matrix and has the potential for detecting norovirus in clinical samples with a small Palmtop Raman spectrometer.