Rapid and reliable multiplex detection of foodborne pathogens is in great demand for ensuring food safety and preventing foodborne diseases. In this study, we developed a highly sensitive SERS immunosensor for the simultaneous detection of multiple foodborne pathogens. Novel biologic interference-free Raman tags synthesized by using the covalent organic frameworks (COF) TBDP as nanocontainer to load biologic interference-free Raman reporters and specific antibodies for interested targets were used to convert and amplify signals of foodborne pathogens. In addition, lectin functionalized magnetic nanoparticles (MNPs@Con A) which could efficiently bind to the carbohydrate constituents on the surface of pathogens were prepared to capture and isolate multiple pathogens simultaneously. The recognition of the target foodborne pathogen impels the generation of sandwich-like composites of MNPs@Con A/pathogen/TBDP@Raman tags, and these composites could be quickly separated from the sample matrix with the assistance of an external magnet. Besides, a mass of Raman reporters was released by eluting the collected MNPs@Con A/pathogen/TBDP@Raman tags composites. Combined with a portable Raman system, characteristic Raman signals (2271 and 2113 cm-1) of the occupied reporters located at the biologic interference-free region were observed and used for the simultaneous detection of two different foodborne pathogenic strains. And an equal limit of detection of 101 CFU/mL was achieved for each strain. This strategy provides new insight into the application of SERS in the detection of pathogenic bacteria.
Keywords: Biologic interference-free; COF; Food safety; Foodborne pathogen; Multiplex detection; SERS immunosensor.
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