Fig. 4: Reporter phage host range analysis and infectivity in human urine and whole blood.

a Plaquing host ranges and bioluminescence detection ranges were determined in growth medium on 52 urological E. coli and E. faecalis isolates and 50 urological Klebsiella spp. isolates from the Zurich Uropathogen Collection (listed in Table S2). Strains were infected with reporter phages and luminescence quantified at 3 h (E. coli and Klebsiella spp.) or 4 h (Enterococcus spp.) post infection. Heat-map shows fold change RLU (FC-RLU). Threshold (TH): ≥ 100 FC-RLU (E. coli and Klebsiella spp.) and 50 FC-RLU (Enterococcus spp.). Clinical isolates that allow for plaque formation are indicated with a green dot. Plaquing host ranges were determined from single spot on the lawn infection assays and bioluminescence detection ranges were from biological quadruplicates (E. coli and K. pneumoniae) or biological triplicates (E. faecalis). b Summary of individual and combined host- and detection ranges. Each reporter phage was able to transduce bioluminescence into strains that were not permissive to plaque formation, effectively leading to an increased detection range compared to the plaquing host range. c Heat maps showing reporter phage-induced luminescence in human urine spiked with 6 selected strains of each bacterial target species. d, e Dose response assay with Enterococcus phages EfS3::nluc and EfS7::nluc in human urine d or blood e spiked with serial dilutions of the clinical isolate E. faecalis Ef24. After 1/10 dilution of the spiked urine/blood with buffered media and a 1 h enrichment, reactions were mixed with 2 × 10⁷ PFU/ml reporter phages and FC-RLU was determined at 2 h and 4 h p.i. FC RLU equals RLU (sample) divided by RLU (phage only control). All data in c–e is shown as mean (±SEM) from biological triplicates. Individual datapoints are shown as black dots. Source data are provided as a Source Data file.