Targeted Single-Phage Isolation Reveals Phage-Dependent Heterogeneous Infection Dynamics

Area of Science:

• Microbiology
• Virology
• Biotechnology

Background:

• Antibiotic resistance poses a critical global health threat, necessitating alternative treatments for multidrug-resistant infections.
• Bacteriophage (phage) therapy, utilizing bacterial viruses, is a promising alternative to antibiotics.
• Isolating effective phages with desirable therapeutic properties and without undesirable traits (e.g., virulence factors) is a significant challenge.

Purpose of the Study:

• To develop a targeted single-phage isolation method for efficient detection and characterization of therapeutic phages.
• To high-throughput characterize phage kinetics and identify phages lacking undesirable bacterial traits.
• To investigate single-cell variations in phage-bacterium interactions during infection.

Main Methods:

• Development of a targeted single-phage isolation technique based on phage similarity to known isolates.
• High-throughput screening for phage kinetics and exclusion of phages carrying bacterial virulence factors.
• Single-cell analysis of phage-infected bacterial populations.

Main Results:

• Successful isolation and characterization of specific phages with therapeutic potential.
• Identification of phages lacking undesirable traits such as bacterial toxins.
• Observation of significant cell-to-cell variations in phage kinetics at the single-cell level, even among infections with the same phage species.

Conclusions:

• The developed method facilitates the isolation and characterization of suitable phages for phage therapy.
• The study highlights the importance of single-cell analysis in understanding phage-bacterium interactions.
• This work contributes to advancing phage therapy as a viable alternative to antibiotics for multidrug-resistant infections.