Rapid Determination of Listeria monocytogenes in Foods Using a Resuscitation/Selection/Kit System Detection

A resuscitation medium consisting of a trypticase soy broth base supplemented with 0.5% yeast extract, 0.25% sodium pyruvate, 0.01 % sodium thiogly col late, and 0.1 % chicken fat was used in the resuscitation of heat-injured and freeze-injured cells of Listeria monocytogenes. After a resuscitation period of 4-h, the medium was made selective through the addition of nalidixic acid, acriflavin, and cycloheximide. The organisms were incubated in the selectivized medium at 35°C for an additional 16 h. The numbers of resuscitated Listeria monocytogenes cells rose from 101 to 107 cells/mL in 20 h. Similar numbers of Staphylococcus aureus, Escherichia coli, and Salmonella bonn were grown together with Listeria monocytogenes; these organisms did not inhibit the growth of Listeria monocytogenes nor interfere with its detection by the Listeria-Tek kit system. The resuscitation/selection/ kit system (RSK) was compared with the methodology in the Bacteriological Analytical Manual (BAM) for the detection of Listeria monocytogenes in 22 naturally contaminated cheese samples: 8 of these were positive by the BAM system and 12 were positive by the RSK system. The 8 Listeria positives found by the BAM system were positive by the RSK system. All 12 Listeria-presumptive positive samples by the RSK system were confirmed to be Listeria monocytogenes. The use of the RSK system enhanced the recovery of the pathogen, and detection was accomplished within 24 h.