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An Optimized Checkerboard Method for Phage-Antibiotic Synergy Detection.

Isidora Nikolic1, Darija Vukovic1, Damir Gavric1

  • 1PK Laboratory, Department of Biology and Ecology, Faculty of Sciences, Trg Dositeja Obradovica 3, University of Novi Sad, 21000 Novi Sad, Serbia.

Viruses
|July 27, 2022
PubMed
Summary

Optimizing the checkerboard method reliably estimates phage-antibiotic synergy. This approach identified synergistic interactions against resistant bacteria, paving the way for personalized phage therapy.

Keywords:
checkerboard methodphage-antibiotic synergytime-kill curve

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Area of Science:

  • Microbiology
  • Bacteriology
  • Antimicrobial Resistance

Background:

  • Phage-antibiotic synergy offers a promising therapeutic strategy against resistant bacteria.
  • Current methods for synergy estimation, like time-kill curve assays, are not suitable for high-throughput screening.
  • A reliable and efficient method is needed to assess phage-antibiotic interactions.

Purpose of the Study:

  • To optimize the checkerboard method for determining phage-antibiotic interactions.
  • To validate the optimized checkerboard method against the gold standard time-kill curve assay.
  • To investigate synergy with simultaneous and successive applications of phages and antibiotics against specific bacterial pathogens.

Main Methods:

  • Optimization of the checkerboard assay for phage-antibiotic synergy.
  • Validation of the checkerboard method using time-kill curve assays.
  • Testing interactions between Pseudomonas phage JG024 and antibiotics (ciprofloxacin, gentamicin, ceftriaxone) against *Pseudomonas aeruginosa*.
  • Testing interactions between Staphylococcus phages MSA6 and SES43300 and antibiotics (ciprofloxacin, gentamicin, oxacillin) against MRSA.

Main Results:

  • The optimized checkerboard method demonstrated reliability and correlated well with time-kill curve assay results.
  • Synergistic interactions were identified between phage JG024 and ciprofloxacin or ceftriaxone against *P. aeruginosa*.
  • Synergy was observed between phage SES43300 and ciprofloxacin against MRSA.
  • Synergy was achieved with simultaneous application, and for *P. aeruginosa*, with a one-hour delay, suggesting simultaneous application is optimal.

Conclusions:

  • The optimized checkerboard method is a reliable tool for assessing phage-antibiotic synergy.
  • This method can facilitate clinical studies and future personalized therapy for multidrug-resistant bacterial infections.
  • Simultaneous application of phages and antibiotics appears to be the most effective strategy for exploiting synergy.