article · Scientific Reports
Multidrug-resistant bacterial infections in the urinary tract require alternative treatment options. Screening fifty multidrug-resistant gram-negative bacterial isolates identified a bacteriophage, named Klebsiella pneumoniae phage KP Ø1, targeting the most prevalent pathogen. The virus demonstrated a fifty percent host range and produced a high lytic titre. Structural analysis revealed an icosahedral head with a short contractile tail, identifying it as a novel Slopekvirus. Genomic sequencing showed a double-stranded DNA genome of 174,591 base pairs with 274 predicted open reading frames. The genome lacks lysogeny-related genes, toxins, and antimicrobial resistance markers, verifying an exclusively lytic lifecycle. Additionally, the phage maintained stability across temperatures from minus twenty to fifty degrees Celsius and within a pH range of seven to nine. These stability characteristics and safety features support further preclinical development of the virus for phage therapy.
The rise of multidrug-resistant bacteria poses severe challenges for treating urinary tract infections when conventional antibiotics fail. Bacteriophages offer a targeted alternative by destroying specific bacteria without harming beneficial microbes. Demonstrating that a newly identified phage is strictly lytic, physically stable, and completely free from harmful resistance genes is an essential milestone in developing safe biological therapies to combat resistant pathogens.
This early-stage research provides a characterised biological candidate for phage therapy against multidrug-resistant urinary tract infections. It could enable biopharmaceutical developers and clinical researchers to formulate targeted antimicrobial treatments or therapeutic cocktails. Because the findings are currently limited to laboratory and genomic characterisation, the virus remains at a preclinical stage requiring extensive in vivo safety testing, clinical trials, and regulatory clearance before reaching healthcare markets.
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The rise of multidrug-resistant (MDR) uropathogenic gram-negative bacteria (GNB) necessitates the development of alternative therapeutic strategies. This study aimed to isolate, phenotypically characterize, and perform whole-genome sequencing of the bacteriophage demonstrating the broadest host range against MDR uropathogens. Fifty MDR GNB isolates were screened for lytic phages. The most promising candidate, Klebsiella pneumoniae phage KP Ø1, was characterized using plaque assay, Transmission Electron Microscopy (TEM), and pH/thermal stability testing. Genomic characterization was performed via whole-genome sequencing (WGS), with functional annotation and lifestyle prediction using PhaBOX and PhageScope software. Klebsiella pneumoniae was the most prevalent MDR uropathogen. Klebsiella pneumoniae phage KP Ø1 exhibited a 50% host range and high lytic titer (10⁸ PFU/mL). TEM revealed an icosahedral head and short contractile tail. Genomic characterization by WGS revealed that Klebsiella pneumoniae phage KP Ø1 possesses a 174,591 bp double-stranded deoxyribonucleic acid (dsDNA) genome containing 274 predicted open reading frames (ORFs). No lysogeny-related genes, toxins, or antibiotic resistance markers were detected, confirming its strictly lytic nature and supporting its potential as a candidate for phage therapy applications. The phage remained stable (10⁸ PFU/mL) across temperatures of - 20 °C to 50 °C; supporting its suitability for long-term biobanking and suggesting potential activity at physiological temperature, and across a pH range of 7-9. Klebsiella pneumoniae phage KP Ø1 is a novel, obligately lytic Slopekvirus whose genomic architecture, stability profile, and absence of lysogeny-associated, virulence, and antimicrobial resistance genes ( AMR) collectively support its candidacy for further preclinical evaluation as a phage therapy agent against uropathogenic MDR Klebsiella pneumoniae.
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DOI: 10.1038/s41598-026-67695-4
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