Recombinant phage probes for Listeria monocytogenes

Monitoring of food and environmental samples for biological threats, such as Listeria monocytogenes, requires probes that specifically bind biological agents and ensure their immediate and efficient detection. There is a need for robust and inexpensive affinity probes as an alternative to antibodies. These probes may be recruited from random peptide libraries displayed on filamentous phage. In this study, we selected from two phage peptide libraries phage clones displaying peptides capable of specific and strong binding to the L. monocytogenes cell surface. The ability of isolated phage clones to interact specifically with L. monocytogenes was demonstrated using enzyme-linked immunosorbent assay ( ELISA) and confirmed by co- precipitation assay. We also assessed the sensitivity of phage - bacteria binding by PCR on phage- captured Listeria cells, which could be detected at a concentration of 104 cells ml(-1). In addition, as proof- of- concept, we tested the possibility of immobilizing the affinity- selected phages to a putative biosensor surface. The quality of phage deposition was monitored by ELISA and fluorescent microscopy. Phage - bacterial binding was confirmed by high power optical phase contrast microscopy. Overall, the results of this work validate the concept of affinity- selected recombinant filamentous phages as probes for detecting and monitoring bacterial agents under any conditions that warrant their recognition, including in food products.