Cefepime-modified magnetic nanoparticles and enzymatic colorimetry for the detection of Listeria monocytogenes in lettuces

A novel sandwich assay for the detection of L. monocytogenes was designed based on antibiotic magnetic separation and enzymatic colorimetry. PEG-mediated cefepime functionalized magnetic nanoparticles (Cefe-PEG-MNPs) was reported for the first time to anchor L. monocytogenes cells with excellent bacterial capture capacity. The capture efficiency of L. monocytogenes in lettuce sample with high concentration (3.1 × 10⁶ CFU/mL) was more than 73.8%. Anti-L. monocytogenes monoclonal antibody was adopted as the second anchoring agent to ensure the specificity for L. monocytogenes, which was co-modified with HRP on the surface of gold nanoparticles (AuNPs-HRP/mAb) to form AuNPs-HRP/mAb@L. monocytogenes@Cefe-PEG-MNPs sandwich complexes, and TMB was added to generate a colorimetric signal. The limit of detection in contaminated lettuce, watermelon juice, and fresh meat samples were both 3.1 × 10² CFU/mL, and the whole assay takes about 110 min. Based on the above facts, the proposed method has great potential for rapid separation and detection of pathogenic bacteria in food.