Plasmon color-preserved gold nanoparticle clusters for high sensitivity detection of SARS-CoV-2 based on lateral flow immunoassay

Lateral flow immunoassays (LFI) have shown great promise for point-of-care (POC) sensing applications, however, its clinical translation is often hindered by insufficient sensitivity for early detection of diseases, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This is mainly due to weak absorption signals of single gold nanoparticles (AuNPs). Here, we developed AuNP clusters that maintain the red color of isolated individual AuNPs, but increase the colorimetric readout to improve the detection sensitivity. The plasmon color-preserved (PLASCOP) AuNP clusters is simply made by mixing streptavidin-coated AuNP core with satellite AuNPs coated with biotinylated antibodies. The biotinylated antibody-streptavidin linker forms a gap size over 15 nm to avoid plasmon coupling between AuNPs, thus maintaining the plasmonic color while increasing the overall light absorption. LFI sensing using PLASCOP AuNP clusters composed of 40 nm AuNPs showed a high detection sensitivity for SARS-CoV-2 nucleocapsid proteins with a limit of detection (LOD) of 0.038 ng mL-1, which was 23.8- and 5.9-times lower value than that of single 15 nm and 40 nm AuNP conjugates, respectively. The PLASCOP AuNP clusters-based LFI sensing also shows good specificity for SARS-CoV-2 nucleocapsid proteins from other influenza and coronaviruses. In a clinical feasibility test, we demonstrated that SARS-CoV-2 particles spiked in human saliva could be detected with an LOD of 54 TCID50 mL-1. The developed PLASCOP AuNP clusters are promising colorimetric sensing reporters that present improved sensitivity in LFI sensing for broad POC sensing applications beyond SARS-CoV-2 detection.