Detection and enumeration of Salmonella enteritidis in homemade ice cream associated with an outbreak: comparison of conventional and real-time PCR methods.
Salmonellosis caused by Salmonella Enteritidis (SE) is a significant cause of foodborne illnesses in the United States. Consumption of undercooked eggs and egg-containing products has been the primary risk factor for the disease. The importance of the bacterial enumeration technique has been enormously stressed because of the quantitative risk analysis of SE in shell eggs. Traditional enumeration methods mainly depend on slow and tedious most-probable-number (MPN) methods. Therefore, specific, sensitive, and rapid methods for SE quantitation are needed to collect sufficient data for risk assessment and food safety policy development. We previously developed a real-time quantitative PCR assay for the direct detection and enumeration of SE and, in this study, applied it to naturally contaminated ice cream samples with and without enrichment. The detection limit of the real-time PCR assay was determined with artificially inoculated ice cream. When applied to the direct detection and quantification of SE in ice cream, the real-time PCR assay was as sensitive as the conventional plate count method in frequency of detection. However, populations of SE derived from real-time quantitative PCR were approximately 1 log higher than provided by MPN and CFU values obtained by conventional culture methods. The detection and enumeration of SE in naturally contaminated ice cream can be completed in 3 h by this real-time PCR method, whereas the cultural enrichment method requires 5 to 7 days. A commercial immunoassay for the specific detection of SE was also included in the study. The real-time PCR assay proved to be a valuable tool that may be useful to the food industry in monitoring its processes to improve product quality and safety.
U.S. Food and Drug Administration, Center for Food Safety and Applied Nutrition, Office of Plant and Dairy Foods, 5100 Paint Branch Parkway, College Park, Maryland 20740, USA. kseo@cfsan.fda.gov
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Seo
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Valentin-Bon
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Brackett
D015169:Colony Count, Microbial / Q000379:methods; D003257:Consumer Product Safety; D004196:Disease Outbreaks; D004531:Eggs / Q000382:microbiology; D005506:Food Contamination / Q000032:analysis*; D006801:Humans; D007054:Ice Cream / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D018570:Risk Assessment; D012478:Salmonella Food Poisoning / Q000453:epidemiology*; D012477:Salmonella enteritidis / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013997:Time Factors