Chimeric Phage Nanoparticles for Rapid Characterization of Bacterial Pathogens: Detection in Complex Biological Samples and Determination of Antibiotic Sensitivity

[Image: see text] Rapid, specific, and sensitive detection of pathogenic bacteria in drink, food, and clinical samples is an important goal for public health. In addition, rapid characterization of antibiotic susceptibility could inform clinical choices and improve antibiotic stewardship. We previously reported a straightforward, inexpensive strategy to detect Gram-negative bacterial pathogens, including Pseudomonas aeruginosa, Vibrio cholerae, and Escherichia coli, taking advantage of the high affinity and specificity of phages for their bacterial hosts. Chimeric phages targeted different bacterial pathogens, and thiolation of the phages induced aggregation of gold nanoparticles (AuNPs), leading to a visible colorimetric response in the presence of at least ∼100 cells of the target bacteria. Here, we apply this strategy to complex biological samples (milk, urine, and swabs from a porcine ex vivo model of P. aeruginosa infection). We also show that this assay can be used to identify the antibiotic susceptibility profile based on detection of bacterial growth in the presence of different antibiotics. The prospect for using phage-conjugated AuNPs to detect bacterial pathogens in clinical samples and guide antibiotic choice is discussed.