Quantitative real-time PCR method for rapid enumeration of Enterobacteriaceae in food

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In food manufacturing companies, the bacteriological examination of foods targets not only foodborne pathogens but total aerobic bacterial counts and hygiene indicator bacteria as well. However, it is hard to establish an examination method using real-time PCR for total aerobic bacteria and hygiene indicator bacteria, since the method needs to target several different bacterial species at the same time. In this study, we developed a rapid detection and quantification method for Enterobacteriaceae using quantitative real-time PCR.First of all, several DNA regions were investigated in order to design PCR primers for specific detection of Enterobacteriaceae. As a result, primers designed within the rplP gene could specifically detect Enterobacteriaceae.Next, the accuracy of the real-time PCR method was confirmed by comparing with a culture method. The difference between these two methods was within one order. These results indicate that the real-timePCR developed in this study could accurately and rapidly quantify Enterobacteriaceae in foods.