Multiplexed detection of Salmonella, E.coli, Campylobacter and Listeria in raw poultry

Authors: Capobianco Jr, Joseph; Armstrong, Cheryl; Chen, Chin Yi; He, Yiping; Counihan, Katrina; Lee, Joseph; Dykes, Gretchen; Johnson, Kathleen; Tilman, Shannon; Koppenhofer, Heather; ATENCIA, JAVIER - Pathotrak Inc; MARTINOS, SEAN - Pathotrak Inc

Submitted to: Foods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/22/2025
Publication Date: 3/25/2025
Citation: Capobianco Jr, J.A., Armstrong, C.M., Chen, C., He, Y., Counihan, K.L., Lee, J., Dykes, G.E., Johnson, K.M., Tilman, S.M., Koppenhofer, H.L., Atencia, J., Martinos, S. 2025. Multiplexed detection of Salmonella, E.coli, Campylobacter and Listeria in raw poultry. Foods. https://doi.org/10.3390/foods14071137.
DOI: https://doi.org/10.3390/foods14071137

Interpretive Summary: Detecting harmful bacteria in food is essential for preventing illnesses. Traditional testing methods can take several days due to long enrichment steps, where bacteria must grow before detection. These processes are not only slow but also require separate steps for different pathogens, increasing complexity. This study tested a faster, enrichment-free method that uses the PathoTrak system to isolate bacteria and the Neogen Molecular Detection System (MDS) to detect multiple pathogens simultaneously. By skipping enrichment, the new method reduced testing time to less than 4 hours for 96 samples. The system successfully identified harmful bacteria like Salmonella, E. coli, Listeria monocytogenes, Listeria species, and Campylobacter in poultry samples, even at very low levels. This innovation promises quicker and more efficient food safety testing, helping regulators and producers respond faster to contamination risks and protect public health.

Technical Abstract: 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, E. coli O157, Listeria monocytogenes, Listeria spp., and Campylobacter in poultry samples, significantly reducing the time to screen samples requiring further evaluation. The system's performance was assessed using inoculated chicken samples at 5 and 25 CFU/g and a range of pathogen 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 Salmonella and Listeria monocytogenes. The system maintained high sensitivity and specificity, achieving perfect agreement for E. coli and Campylobacter. This study highlights the potential of enrichment-free, multiplex detection to streamline food safety testing by reducing time-to-results, enhancing efficiency, and providing reliable pathogen quantification across multiple targets.