Specific separation and sensitive detection of foodborne pathogens by phage-derived bacterial-binding protein-nano magnetic beads coupled with smartphone-assisted paper sensor

Foodborne pathogen infection poses a significant threat to public health and is considered as one of the most serious hazards in global food safety. Herein, a sensitive and efficient method for on-site monitoring of foodborne pathogens was developed by using a smartphone-assisted paper-sensor combined with phage-derived bacterial-binding proteins-nano magnetic beads (PBPs-MBs). PBPs including tail fiber protein (TFP:gp13), cell-wall binding domain (CBD) of endolysin and tailspike protein (TSP) coated on the surface of MBs were applied for rapid separation and enrichment of targeted bacteria (Escherichia coli O157:H7, Staphylococcus aureus and Salmonella typhimurium, respectively) from food samples in 20 min before detection on paper-based sensors. The paper-based sensor was loaded with the lytic agent (polymyxin B) to induce bacterial lysis and release specific endogenous enzymes. Subsequently, three distinct chromogenic substrates were hydrolyzed by their corresponding enzymes, resulting in characteristic color changes on the paper, respectively. In addition, a smartphone APP for red-green-blue (RGB) color analysis of paper was able to directly detect three foodborne pathogens. As a result, the limit of detection (LOD) values for three foodborne pathogens were found to be 2.44 × 10