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Phage-Host Interaction Analysis by Flow Cytometry Allows for Rapid and Efficient Screening of Phages
Luís D R Melo1,2, Rodrigo Monteiro1,2, Diana P Pires1,2
1LIBRO-Laboratório de Investigação em Biofilmes Rosário Oliveira, Centre of Biological Engineering, Campus de Gualtar, University of Minho, 4700-057 Braga, Portugal.
Antibiotics (Basel, Switzerland)
|February 25, 2022
Summary
Flow cytometry offers a rapid method to screen phages for bacterial infection. This technique quickly assesses phage susceptibility, crucial for selecting effective phage therapy treatments.
Area of Science:
- Microbiology
- Biotechnology
- Molecular Biology
Background:
- Phage therapy is a promising alternative to antibiotics, necessitating efficient screening methods.
- Current phage screening methods are time-consuming, hindering rapid clinical application.
Purpose of the Study:
- To establish flow cytometry (FC) as a rapid and reliable method for assessing bacterial susceptibility to phage infection.
- To evaluate FC for high-throughput, real-time screening of phages against specific bacterial hosts.
Main Methods:
- Characterized phage-bacteria interactions using flow cytometry (FC) with SYTO BC and PI staining.
- Performed synchronous infection assays and gene expression analysis via qPCR.
- Correlated FC and qPCR data with phage propagation assays.
Main Results:
- Flow cytometry detected increased SYTO BC median fluorescence intensity (MFI) within 25 minutes of phage infection, correlating with phage gene expression.
- This increase in MFI indicates successful phage gene expression and host susceptibility.
- The method effectively distinguished true infection from lysis from without, avoiding false positives.
Conclusions:
- Flow cytometry provides a rapid, reliable, and high-throughput method for screening phage efficacy against bacterial hosts.
- This FC-based approach can significantly accelerate phage selection for clinical phage therapy applications.
