Multifunctional Nanoprobe-Amplified Enzyme-Linked Immunosorbent Assay on Capillary: A Universal Platform for Simple, Rapid, and Ultrasensitive Dual-Mode Pathogen Detection

Although the traditional enzyme-linked immunosorbent assay (ELISA) has been widely applied in pathogen detection and clinical diagnostics, it always suffers from complex procedures, a long incubation time, unsatisfying sensitivity, and a single signal readout. Here, we developed a simple, rapid, and ultrasensitive platform for dual-mode pathogen detection based on a multifunctional nanoprobe integrated with a capillary ELISA (CLISA) platform. The novel capture antibodies-modified capillaries can act as a swab to combine in situ trace sampling and detection procedures, eliminating the dissociation between sampling and detection in traditional ELISA assays. With excellent photothermal and peroxidase-like activity, the Fe₃O₄@MoS₂ nanoprobe with a unique p-n heterojunction was chosen as an enzyme substitute and amplified signal tag to label the detection antibody for further sandwich immune sensing. As the analyte concentration increased, the Fe₃O₄@MoS₂ probe could generate dual-mode signals, including remarkable color changes from the chromogenic substrate oxidation as well as photothermal enhancement. Moreover, to avoid false negative results, the excellent magnetic capability of the Fe₃O₄@MoS₂ probe can be used to pre-enrich the trace analytes, amplifying the detection signal and enhancing the immunoassay's sensitivity. Under optimal conditions, specific and rapid detection of SARS-CoV-2 has been realized successfully based on this integrated nanoprobe-enhanced CLISA platform. The detection limits were 5.41 pg·mL^-1 for the photothermal assay and 150 pg·mL^-1 for the visual colorimetric assay. More importantly, the simple, affordable, and portable platform can also be expanded to rapidly detect other targets such as Staphylococcus aureus and Salmonella typhimurium in practical samples, making it a universal and attractive tool for multiple pathogen analysis and clinical testing in the post COVID-19 era.