Simultaneous detection of Salmonella enterica, Escherichia coli and Listeria monocytogenes in food using a light scattering sensor.

Abstract

To investigate the use of a light scattering sensor, BActerial Rapid Detection using Optical scattering Technology (BARDOT) coupled with a multipathogen selective medium, Salmonella, Escherichia and Listeria (SEL), for concurrent detection of the three major foodborne pathogens in a single assay. BARDOT was used to detect and distinguish the three major pathogens, Salmonella enterica, Shiga toxin-producing Escherichia coli (STEC) and Listeria monocytogenes from food based on colony scatter signature patterns on SEL agar (SELA). Multiple strains of three test pathogens were grown on SELA, and BARDOT was used to generate colony scatter image libraries for inclusive (SEL Library) and exclusive (non-SEL Library) bacterial group. These pathogens were further differentiated using the SEL scatter image library. Raw chicken and hotdog samples were artificially inoculated with pathogens (100 CFU per 25g each), and enriched in SEL broth at 37°C for 18h and colonies were grown on SELA for 11-22h before screening with BARDOT. The BARDOT sensor successfully detected and differentiated Salmonella, STEC and Listeria on SELA with high classification accuracy 92-98%, 91-98% and 83-98% positive predictive values (PPV) respectively; whereas the nontarget strains showed only 0-13% PPV. BARDOT-identified colonies were further confirmed by multiplex PCR targeting inlB gene of L. monocytogenes, stx2 of STEC and sefA of S. enterica serovar Enteritidis. The results show that BARDOT coupled with SELA can efficiently screen for the presence of three major pathogens simultaneously in a test sample within 29-40h. This innovative SELA-BARDOT detection platform can reduce turnaround time and economic burden on food industries by offering a label-free, noninvasive on-plate multipathogen screening technology for reducing microbial food safety and public health concerns.