Pomegranate-Inspired Silica Nanotags Enable Sensitive Dual-Modal Detection of Rabies Virus Nucleoprotein

The outbreak of rabies virus (RABV) in Asia and Africa has attracted widespread concern due to its 100% mortality rate, and RABV detection is crucial to its diagnosis and treatment. Herein, we report a sensitive and reliable strategy for the dual-modal RABV detection using pomegranate-shaped dendritic silica nanospheres fabricated with densely incorporated quantum dots (QDs) and horseradish peroxidase (HRP)-labeled antibody. The immunoassay involves the specific interaction between virus and nanospheres-conjugated antibody coupled with robust fluorescence signal originating from QDs and naked-eye discernible colorimetric signal on the oxTMB. The ultrahigh loading capacity of QDs enables the detection limit down to 8 pg/mL via fluorescence modality, a 348-fold improvement as compared with conventional enzyme-linked immunosorbent assay (ELISA). In addition, the detection range was from 1.20 × 10² to 2.34 × 10⁴ pg/mL by plotting the absorbance at 652 nm with RABV concentrations with a detection limit of 91 pg/mL, which is nearly 2 order of magnitude lower than that of the conventional ELISA. Validated with 12 brain tissue samples, our immunoassay results are completely consistent with polymerase chain reaction (PCR) results. Compared with the PCR assay, our approach requires no complex sample pretreatments or expensive instruments. This is the first report on RABV diagnosis using nanomaterials for colorimetry-based prescreening and fluorescence-based quantitative detection, which may pave the way for virus-related disease diagnosis and clinical analysis.