Rapid and Quantitative Detection of <I>Escherichia coli</I> and <I>Enterococcus</I> in Environmental Waters Using Real-Time PCR

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  • OGURA Yudai
    Advanced Course of Civil and Environmental Engineering, Hachinohe National College of Technology
  • YAGUCHI Junichi
    Department of Civil and Environmental Engineering, Hachinohe National College of Technology
  • KOMATSU Toshiya
    Department of Civil and Environmental Engineering, Nagaoka University of Technology

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Other Title
  • リアルタイムPCRによる水環境水中の糞便汚染指標細菌の定量
  • リアルタイム PCR ニ ヨル ミズ カンキョウ スイチュウ ノ フンベン オセン シヒョウ サイキン ノ テイリョウ
  • Rapid and Quantitative Detection of Escherichia coli and Enterococcus in Environmental Waters Using Real-Time PCR

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A rapid and quantitative polymerase chain reaction (PCR) method for determining the numbers of two fecal pollution indicators, E. coli and Enterococcus was developed. The quantitative detection of E. coli and Enterococcus was performed on the basis of the uidA and 23S-rRNA gene sequences, respectively. A linear relationship was observed between the log concentration of each gene and the threshold cycle value obtained from the PCR amplification curve. Using these standard PCR curves, the concentrations of E. coli and Enterococcus were measured in environmental water samples collected from a wastewater treatment plant and four bodies of water located in the northern part of Japan. Measurements by the real-time PCR method were compared with E. coli and Enterococcus counts determined by cultivation methods such as membrane filter and MPN analyses. The PCR method gave quicker results and higher E. coli and Enterococcus counts than the traditional cultivation methods. These higher counts suggested the ability of the PCR method to detect DNA from both culturable and nonculturable or dead bacteria. Regression analysis of these measurements showed significant positive correlations between the PCR method and the traditional cultivation methods with correlation coefficients of over 0.7.

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