There is a need for improved screening methods of bacterial contaminants in food and clinical samples. Bacteriophages are ideal detection elements due to their host specificity, short amplification time, ability to differentiate between viable and non-viable cells, and capability to delivery genetic material. As a proof-of-principal, we have genetically engineering bacteriophage T7 to enable rapid detection of <i>E. coli</i> in a water sample. We have modified T7 to carry an enzymatic reporter, a His-tagged TEV protease, for overexpression in <i>E. coli</i>. The His-tag allows for TEV separation and concentration with magnetic beads. The separated TEV is added to a solution containing a peptide composed of identical subunits that are cleaved when TEV is present. MALDI-TOF is used to rapidly screen this sample for changes in peptide composition. This scheme will allow for rapid identification of potentially contaminated samples for corrective response and further analysis.

Bacteriophage-based scheme to enable rapid MALDI-TOF screening of samples for bacterial contaminants

249th National Meeting (2015)

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