Rapid separation and detection of Listeria monocytogenes with the combination of phage tail fiber protein and vancomycin-magnetic nanozyme

In this work, a lateral flow assay for Listeria monocytogenes was developed based on phage tail fiber protein (TFP) and triple-functional nanozyme probes with capture-separation-catalytic activity. Inspired by interaction between phage and bacteria, TFP of L. monocytogenes phage was immobilized on test line as capture molecule, which replaced traditional antibody and aptamer. After Gram-positive bacteria was captured and separated from samples by nanozyme probes modified with vancomycin (Van), TFP specifically recognized L. monocytogenes and overcame non-specific binding of Van. Special color reaction between Coomassie Brilliant Blue and bovine serum albumin which was an amplification carrier on probe was simply utilized as control zone to replace traditional control line. Relying on enzyme-like catalytic activity of nanozyme, this biosensor realized improved sensitivity and colorimetric quantitative detection with a detection limit of 10 CFU mL-1. Analytic performance results suggested this TFP-based biosensor provided a portable, sensitive and specific strategy to detect pathogen.