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 Salmonella, Escherichia coli O157, Listeria monocytogenes, Listeria spp., and Campylobacter 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 (10(2)-10(8) CFU/mL). The MDS system demonstrated robust detection for most pathogens, with strong correlations between theoretical inoculation levels and MDS-calculated concentrations (R(2) > 0.85 for all pathogens). However, detection variability was observed at lower concentrations for Salmonella and Listeria monocytogenes. The system maintained high sensitivity and specificity, achieving a Cohen's Kappa coefficient for E. coli and Campylobacter. 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.