Bacteriophage replication on permissive host cells in fused silica capillary with nanostructured part as potential of electrophoretic methods for developing phage applications

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Authors

HORKÁ Marie KARÁSEK Pavel ROTH Michal ŠTVERÁKOVÁ Dana ŠALPLACHTA Jiří RŮŽIČKA Filip PANTŮČEK Roman

Year of publication 2021
Type Article in Periodical
Magazine / Source Talanta
MU Faculty or unit

Faculty of Science

Citation
Web https://doi.org/10.1016/j.talanta.2020.121800
Doi http://dx.doi.org/10.1016/j.talanta.2020.121800
Keywords Capillary electrophoretic methods; Fused silica capillary with roughened part; Microliter sample volumes; Dynamic adhesion of bacterial cells; Staphylococcus aureus; Bacteriophage propagation
Description 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.
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