Multiplexed Detection of Salmonella, Escherichia coli, Campylobacter, and Listeria in Raw Poultry

The detection of foodborne pathogens is a critical aspect of ensuring food safety. Traditional methods rely on time-intensive enrichment steps and pathogen-specific assays, extending testing timelines and limiting throughput. This study evaluates an enrichment-free, multiplexed pathogen detection workflow combining the Pathotrak system for bacterial separation and the Neogen Molecular Detection System (MDS) for detection. The workflow enables simultaneous detection of <i>Salmonella</i>, <i>Escherichia coli</i> O157, <i>Listeria monocytogenes</i>, <i>Listeria</i> spp., and <i>Campylobacter</i> in poultry samples, significantly reducing the time taken to screen samples requiring further evaluation. The system's performance was assessed using inoculated chicken samples over a range of bacterial concentrations (10²-10⁸ CFU/mL). The MDS system demonstrated robust detection for most pathogens, with strong correlations between theoretical inoculation levels and MDS-calculated concentrations (R² > 0.85 for all pathogens). However, detection variability was observed at lower concentrations for <i>Salmonella</i> and <i>Listeria monocytogenes</i>. The system maintained high sensitivity and specificity, achieving a Cohen's Kappa coefficient for <i>E. coli</i> and <i>Campylobacter</i>. This study highlights the potential of enrichment-free, multiplex detection to streamline food safety testing by reducing the time to results, enhancing efficiency, and providing reliable pathogen quantification across multiple targets.