Fig. 5: Field evaluation demonstrates rapid, sensitive, and reliable detection of E. coli, Enterococcus spp. and Klebsiella spp. in patient urine.

Workflow of the urine diagnostic assay is shown in a. Fresh patient urine was collected and bacterial metabolic activity activated by a short 1 h enrichment in growth media followed by addition of reporter phages. Luminescence was quantified at 3 h (E. coli, Klebsiella spp.) and 4 h (Enterococcus spp.) post infection. b Luminescence quantification (RLU) was correlated to bacterial CFU/ml counts from differential/selective plating of 206 patient urine samples on UriSelect and KFS agar plates. In parallel, all results were confirmed by the diagnostic laboratory of the Institute of Medical Microbiology (IMM). Urine samples were derived from two different hospitals (Site 1 and Site 2). Thresholds (TH) for luminescence were ≥ 100 FC-RLU (E. coli, green and Klebsiella spp., purple) and 7.5 FC-RLU (Enterococcus spp., blue). The test was considered positive if at least one phage per target pathogen had activity >TH. False negatives (FN, red) are samples with no luminescence but bacterial growth ( ≥ 10³ CFU/ml), false positives (FP, yellow) exhibit luminescence but no bacterial growth. c Summary of the test performance showing calculated sensitivity, specificity, accuracy for all target pathogens at both sampling sites. Some elements of the figure were created using biorender.com. Strains from the field evaluation are listed in Table S3. Source data are provided as a Source Data file.