High-throughput assay for temporal kinetic analysis of lytic coliphage activity.
Plaque analysis allows the determination of phage titer and multiplicity of infection. Yet, this overnight assay provides only endpoint results, ignoring kinetic aspects. We introduce an alternative high-throughput and rapid method for kinetic analysis of lytic coliphage activity. Escherichia coli was infected with serial dilutions of MS2 coliphage, and bacterial growth was monitored using a multi-well plate reader providing within hours the equivalent data as obtained overnight. Additional information is yielded, including phage replication rate, progeny size per cycle, and viral propagation during bacterial growth. This method offers further insights into physicochemical mechanisms of lytic coliphage infection and temporal control. It also provides a virus-host interaction acumen.
Department of Molecular Microbiology and Biotechnology, Tel Aviv University, Tel Aviv 69978, Israel.
Dan
D
Davidi
Dorin
D
Sade
Shlomit
S
Schuchalter
Ehud
E
Gazit
D004926:Escherichia coli / Q000254:growth & development / Q000821:virology*; D057166:High-Throughput Screening Assays; D007700:Kinetics; D017909:Levivirus / Q000254:growth & development / Q000472:pathogenicity / Q000502:physiology*; D013997:Time Factors
Burst size; Lytic coliphages; Phage infection; Plaque assay; Replication rate