An assay to identify the common food-borne pathogens Salmonella, Escherichia coli, Shigella, and Listeria monocytogenes was developed in collaboration with Ibis Biosciences (a division of Abbott Molecular) for the Plex-ID biosensor system, a platform that uses electrospray ionization mass spectroscopy (ESI-MS) to detect the base composition of short PCR amplicons. The new food-borne pathogen (FBP) plate has been experimentally designed using four gene segments for a total of eight amplicon targets. Initial work built a DNA base count database that contains more than 140 Salmonella enterica, 139 E. coli, 11 Shigella, and 36 Listeria patterns and 18 other Enterobacteriaceae organisms. This assay was tested to determine the scope of the assay's ability to detect and differentiate the enteric pathogens and to improve the reference database associated with the assay. More than 800 bacterial isolates of S. enterica, E. coli, and Shigella species were analyzed. Overall, 100% of S. enterica, 99% of E. coli, and 73% of Shigella spp. were detected using this assay. The assay was also able to identify 30% of the S. enterica serovars to the serovar level. To further characterize the assay, spiked food matrices and food samples collected during regulatory field work were also studied. While analysis of preenrichment media was inconsistent, identification of S. enterica from selective enrichment media resulted in serovar-level identifications for 8 of 10 regulatory samples. The results of this study suggest that this high-throughput method may be useful in clinical and regulatory laboratories testing for these pathogens.
Mass spectrometry is an established analytical technique with growing applications within microbiology. With high sensitivity and high resolution, mass spectrometry can be used to differentiate microbial species based on subcellular variations. Recently, several articles concerning the application of either matrix-assisted laser desorption ionization (MALDI) (6, 14, 36) or electrospray ionization (ESI) mass spectrometry (MS) (13,21,30,37) to the detection and identification of microbes have been published. While some methods examine protein expression, this work centers on the use of nucleic acid information to identify bacteria.MS techniques involving the analysis of DNA take advantage of the difference in mass between strands with different base compositions. In order to utilize MS for DNA-based identification of bacteria, a region of DNA that varies between species or subspecies is amplified by PCR, and the mass of this amplicon is then determined. Since the exact masses of the individual bases in DNA are known, the quantity of each of these bases within the amplified sequence can be calculated based on the exact mass of the strand. While the exact sequence is not obtained through this method, the base compositions, or base counts, can provide enough information to discriminate between species, subspecies, and even serovars depending on the organism and the assay (15,19).This technique is comparable to oth...