1 research outputs found
Bacterial Detection Using Unlabeled Phage Amplification and Mass Spectrometry through Structural and Nonstructural Phage Markers
According to the World Health Organization,
food safety is an essential
public health priority. In this context, we report a relevant proof
of feasibility for the indirect specific detection of bacteria in
food samples using unlabeled phage amplification coupled to ESI mass
spectrometry analysis and illustrated with the model phage systems
T4 and SPP1. High-resolving power mass spectrometry analysis (including
bottom-up and top-down protein analysis) was used for the discovery
of specific markers of phage infection. Structural components of the
viral particle and nonstructural proteins encoded by the phage genome
were identified. Then, targeted detection of these markers was performed
on a triple quadrupole mass spectrometer operating in the selected
reaction monitoring mode. <i>E. coli</i> at 1 Ă— 10<sup>5</sup>, 5 Ă— 10<sup>5</sup>, and 1 Ă— 10<sup>6</sup> CFU/mL
concentrations was successfully detected after only a 2 h infection
time by monitoring phage T4 structural markers in Luria–Bertani
broth, orange juice, and French bean stew (“cassoulet”)
matrices. Reproducible detection of nonstructural markers was also
demonstrated, particularly when a high titer of input phages was required
to achieve successful amplification. This strategy provides a highly
time-effective and sensitive assay for bacterial detection