Monitoring phage infection and lysis of surface-immobilized bacteria by QCM-D

<jats:title>Abstract</jats:title> <jats:p>While being a promising approach for the treatment of infections caused by drug-resistant, pathogenic bacteria, the clinical implementation of phage therapy still faces several challenges. One of these challenges lies in the high strain-specificity of most bacteriophages, which makes it necessary to screen large phage collections against the target pathogens in order to identify suitable candidates for the formulations of personalized therapeutic phage cocktails. In this work, we evaluate the potential of quartz crystal microbalance with dissipation monitoring (QCM-D) to identify and detect phage infection and subsequent lysis of bacteria immobilized on the surfaces of the QCM-D sensors. Using lytic <jats:italic>Escherichia coli</jats:italic> phage T7 as a model, we show that phage infection of <jats:italic>E. coli</jats:italic> cells results in various unique alterations in the behaviors of the frequency (Δ<jats:italic>f</jats:italic>) and dissipation (Δ<jats:italic>D</jats:italic>) signals, which are not observed during exposure of the <jats:italic>E. coli</jats:italic> strain to non-infectious <jats:italic>Bacillus subtilis</jats:italic> phage phi29 at similar concentration. To aid future phage screening campaigns, we furthermore identify a single measurement parameter, i.e., the spread between the different overtones of Δ<jats:italic>D</jats:italic>, that can be used to detect phage-induced lysis. For T7 infection of <jats:italic>E. coli</jats:italic>, this is achieved within 4 h after inoculation, including immobilization and growth of the bacteria on the sensor surface, as well as the completed phage propagation cycle. Given the commercial availability of highly automated multichannel systems and the fact that this approach does not require any sensor modifications, QCM-D has the potential to become a valuable tool for screening medium-sized phage collections against target pathogens.</jats:p> <jats:p> <jats:bold>Graphical Abstract</jats:bold> </jats:p>