Detection of Phage’s Lytic Activity Against Carbapenemase-Producing Klebsiella pneumoniae Isolates Using a High-Throughput Microbroth Growth Inhibition Assay DOI Creative Commons
Paschalis Paranos, Spyros Pournaras, Joseph Meletiadis

et al.

Infectious Diseases and Therapy, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 22, 2024

The host range of phages is usually assessed with the agar overlay method. However, this method both cumbersome and subjective. Therefore, a microbroth assay was developed to assess lytic activity patterns in against collection carbapenemase-producing Klebsiella pneumoniae (CRKP) isolates. 11 K. pneumoniae-specific 8 non-repetitive well-characterized CRKP isolates by monitoring optical density (OD) at 630 nm for 24 h different phage concentrations (5 × 109–5 103 PFU/ml) two bacterial inocula 106 5 108 CFU/ml). phage–bacteria pairs transparent/semi-transparent (N = 7), turbid 6), no 6) lysis compared statistically growth inhibition 6 analysis variance (ANOVA), receiver operating characteristic curves (ROC) Fisher's exact test. Optimal cutoffs were determined, sensitivity specificity calculated. Statistically significant differences ≥ PFU/ml found between transparent/semi-transparent, turbid, lysis. ROC curve indicated an optimal cutoff 31% high bacteria detecting 61% high-phage low-bacteria sensitivity/specificity 100%/100% 100%/86%, respectively. provided fast, reliable, objective results host-range differentiating high-throughput format.

Language: Английский

Revisiting therapeutic options against Resistant Klebsiella Pneumoniae infection: phage therapy is key DOI
Jiabao Xing,

Rong-jia Han,

Jinxin Zhao

et al.

Microbiological Research, Journal Year: 2025, Volume and Issue: unknown, P. 128083 - 128083

Published: Jan. 1, 2025

Language: Английский

Citations

1
In Vitro Interactions Between Bacteriophages and Antibacterial Agents of Various Classes Against Multidrug-Resistant Metallo-β-Lactamase-Producing Pseudomonas aeruginosa Clinical Isolates DOI Creative Commons
Paschalis Paranos, Sophia Vourli, Spyros Pournaras

et al.

Pharmaceuticals, Journal Year: 2025, Volume and Issue: 18(3), P. 343 - 343

Published: Feb. 27, 2025

Background: Combination therapy with antibiotics and phages has been suggested to increase the antibacterial activity of both phages. We tested in vitro five belonging different classes combination lytic bacteriophages against multidrug-resistant metallo-β-lactamase (MBL)-producing Pseudomonas aeruginosa isolates. Material/Methods: A total 10 non-repetitive well-characterized MBL-producing P. isolates (5 NDM, 5 VIM) co-resistant aminoglycosides quinolones were used. Phage–antibiotic interactions assessed using an ISO-20776-based broth microdilution checkerboard assay 96-well microtitration plates. Two-fold dilutions colistin (8–0.125 mg/L), ciprofloxacin, meropenem, aztreonam, amikacin (256–4 mg/L) combined ten-fold × 109–5 100 PFU/mL) Pakpunavirus, Phikzvirus, Pbunavirus, Phikmvvirus genus. Plates incubated at 35 ± 2 °C for 24 h, minimum inhibitory concentration (MICA) (MICP) determined as lowest drug phage concentration, resulting <10% growth based on photometric reading 550 nm. Interactions fractional index (FICi) three independent replicates clinical relevance reversal phenotypic resistance. The statistical significance each alone was GraphPad Prism 8.0. Results: Synergistic additive found 60–80% all drugs. FICis statistically significantly lower than 0.5 (p = 0.005), ciprofloxacin 0.02), meropenem 0.003), 0.002). clinically achievable concentrations colistin, amikacin, a resistance observed most strains (63–64%) meropenem. Antagonism few tested. Phage vB_PaerM_AttikonH10 vB_PaerP_AttikonH4 Phikzvirus genus, respectively, showed either synergistic (FICi ≤ 0.35) or effects Conclusions: Synergy drugs showing strong synergy Taking into account wide utility jumbo obtained, findings can enhance currently ineffective

Language: Английский

Citations

0
The global resistance problem and the clinical antibacterial pipeline DOI
Ursula Theuretzbacher

Nature Reviews Microbiology, Journal Year: 2025, Volume and Issue: unknown

Published: April 10, 2025

Language: Английский

Citations

0
Detection of Phage’s Lytic Activity Against Carbapenemase-Producing Klebsiella pneumoniae Isolates Using a High-Throughput Microbroth Growth Inhibition Assay DOI Creative Commons
Paschalis Paranos, Spyros Pournaras, Joseph Meletiadis

et al.

Infectious Diseases and Therapy, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 22, 2024

The host range of phages is usually assessed with the agar overlay method. However, this method both cumbersome and subjective. Therefore, a microbroth assay was developed to assess lytic activity patterns in against collection carbapenemase-producing Klebsiella pneumoniae (CRKP) isolates. 11 K. pneumoniae-specific 8 non-repetitive well-characterized CRKP isolates by monitoring optical density (OD) at 630 nm for 24 h different phage concentrations (5 × 109–5 103 PFU/ml) two bacterial inocula 106 5 108 CFU/ml). phage–bacteria pairs transparent/semi-transparent (N = 7), turbid 6), no 6) lysis compared statistically growth inhibition 6 analysis variance (ANOVA), receiver operating characteristic curves (ROC) Fisher's exact test. Optimal cutoffs were determined, sensitivity specificity calculated. Statistically significant differences ≥ PFU/ml found between transparent/semi-transparent, turbid, lysis. ROC curve indicated an optimal cutoff 31% high bacteria detecting 61% high-phage low-bacteria sensitivity/specificity 100%/100% 100%/86%, respectively. provided fast, reliable, objective results host-range differentiating high-throughput format.

Language: Английский

Citations

1