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

Abstract

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 <i>SARS-CoV-2</i> 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 <i>Staphylococcus aureus</i> and <i>Salmonella typhimurium</i> in practical samples, making it a universal and attractive tool for multiple pathogen analysis and clinical testing in the post COVID-19 era.