Real-time monitoring by interferometric light microscopy of phage suspensions for personalised phage therapy

Phage therapy uses viruses (phages) against antibiotic resistance. Tailoring treatments to specific patient strains requires stocks of various highly concentrated purified phages. It, therefore, faces challenges: titration duration and specificity to a phage/bacteria couple; purification affecting stability; and highly concentrated suspensions tending to aggregate. To address these challenges, interferometric light microscopy (ILM), characterising particles (size, concentration, and visual homogeneity) within minutes, was applied herein to anti-Staphylococcus aureus myovirus phage suspensions. Particle concentration was linearly correlated with phage infectious titre (R² > 0.97, slope: 3 particles/plaque forming units (PFU)) at various degrees of purification, allowing to approximate the infectious titre for suspensions ≥ 3 × 10⁸ PFU/mL, thereby encompassing most therapeutic doses. Purification narrowed and homogenised particle distribution while maintaining therapeutic concentrations. When compared to dynamic light scattering, electrophoretic mobility, and UV/Visible-spectroscopy, ILM best detected aggregates according to our homemade scoring. Although ILM has certain limitations, such as the inability to detect podoviruses (hydrodynamic diameter < 80 nm), or to measure particles in low-concentrated suspensions (< 10⁸ particles/mL), the present proof-of-concept positions this technique as a valuable quality control tool, as a complement to titration rather than a replacement for this technique, for phage suspensions, paving the way for further investigations.