Antimicrobial peptide-based magnetic separation combined with multiplex PCR for the simultaneous detection of multiple foodborne pathogens in food samples

A broad-spectrum bacterial enrichment strategy was developed using antimicrobial peptide-modified magnetic beads (MBs@AMP). • The method combined with mPCR simultaneously detects five foodborne pathogens in fruit juice samples. • This approach eliminates food matrix interference and improves detection sensitivity compared to conventional PCR. Rapid detection of foodborne pathogens in food samples is essential to mitigate threats to public health and ensure food safety. Due to the complexity of food matrix and low concentration of pathogens, traditional detection methods have certain limitations in actual detection. The effective enrichment of foodborne pathogens during sample pretreatment will greatly improve the sensitivity and accuracy of subsequent detection. In this study, an antimicrobial peptide (AMP) was designed as a capture element capable of binding multiple bacterial species, including both Gram-negative and Gram-positive ones. The AMP was loaded on the surface of the magnetic beads (MBs@AMP) for magnetic separation. The MBs@AMP were then combined with multiplex PCR (MBs@AMP-mPCR) to rapidly and sensitively detect Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Shigella spp. , and Staphylococcus aureus in food samples. The detection limit of MBs@AMP-mPCR was 100-fold lower than that of conventional PCR, which is of great significance for the rapid detection of foodborne pathogens.