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Application of Novel PCR Method to Microbial Monitoring in Food Processing and Safety

Published by the American Society of Agricultural and Biological Engineers, St. Joseph, Michigan www.asabe.org

Citation:  Paper number  036172,  2003 ASAE Annual Meeting . (doi: 10.13031/2013.14150) @2003
Authors:   T. KIMURA, H. KOYAMA, N. SHIMIZU, T. INOUE
Keywords:   fermented food, SSC-PCR, food safety, microbial monitoring, process control, yogurt

An effective and rapid monitoring method that can detect various bacteria for ensuring safety of food and for proper control of fermentation process is likely to be a most important tool in food industry. In this study, the Novel PCR method named SSC (Single strain counting)-PCR method which can detect a wide range of bacteria (Kimura et al, ASAE Paper, 2001) was applied to a fermented food model in order to control microbial activity in the mixed culture state fermentation or to identify harmful species from food in the ambient environment.

In our preliminary experiment, DNA extraction was improved by introduction of stronger enzyme (Proteinase K) different from our previous one. We could extract more volume of DNA from the sample, which contributed to a stability of the SSC-PCRs performance. We carried out two types of experiment. Yogurt that is produced by two species of lactic acid bacteria was used as a simple model in the experiments.

In the first experiment, the SSC-PCR could clearly discriminate the two species with use of 48 selected primers. The SSC-PCR could also identify two foreign micro-organisms (E. coli., and B. sub.) from yogurt with use of 8 selected primers in the second experiment. From these results, the SSC-PCR can be used as a tool for rapid monitoring of complexed microflora in food, but the sensitivity is not yet enough to be a practical monitoring method for the samples having lower density of microbial population.

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