Bacteriophage replication on permissive host cells in fused silica capillary with nanostructured part as potential of electrophoretic methods for developing phage applications
HORKÁ, Marie, Pavel KARÁSEK, Michal ROTH, Dana ŠTVERÁKOVÁ, Jiří ŠALPLACHTA et. al.
Bacteriophage replication on permissive host cells in fused silica capillary with nanostructured part as potential of electrophoretic methods for developing phage applications
Authors
HORKÁ, Marie, Pavel KARÁSEK, Michal ROTH, Dana ŠTVERÁKOVÁ (203 Czech Republic, belonging to the institution), Jiří ŠALPLACHTA, Filip RŮŽIČKA (203 Czech Republic, belonging to the institution) and Roman PANTŮČEK (203 Czech Republic, guarantor, belonging to the institution)
Phage therapy could offer a safe and effective alternative to antibiotic treatment of infections caused by Gram-positive bacterium Staphylococcus aureus that have emerged as a significant threat in hospital and community environment and is attracting growing interest among clinicians. The legislation process of approving the phage therapeutics by pharmaceutical authorities requires rapid analytical techniques for assessment of phage activity. Here, we present a three-step method for on-line monitoring the phage effect on bacterial cells dynamically adhered from microliter volumes of high conductivity matrix onto the inner surface of fused silica capillary with a part etched with supercritical water. Phage K1/420 particles of the Kayvirus genus generated by propagation on the host S. aureus cells together with the uninfected cells were concentrated, separated and detected using capillary electrophoretic methods. The phage interactions with selected S. aureus strains exhibiting differences in phage susceptibility were compared. The method allowed determination of the phage burst size and time of phage latent period in analyzed strains. Apart from enumeration of bacteriophages by the plaque assays, the proposed method is suitable for phage activity testing.
Links
GA18-13064S, research and development project
Name: Analýza interakcí mezi polyvalentním terapeutickým fágem druhu Twort-like a jeho hostitelem Staphylococcus aureus
Investor: Czech Science Foundation
NV16-29916A, research and development project
Name: Využití bakteriofágů v léčbě nozokomiálních infekcí spojených s multirezistencí či tvorbou biofilmu
VI20172020069, research and development project
Name: Vývoj moderních instrumentálních metod pro rychlou detekci a identifikaci vybraných B-agens a toxinů (Acronym: VMIMRDIBT)