Molecular monitoring of Escherichia coli O157: H7 sterilization rate using qPCR and propidium monoazide treatment

Authors


Correspondence

Yu Yi-gang, Department Light Industry and Food Sciences, South China University of Technology, Guangzhou City, Guangdong Province 510640, China.

E-mail: yuyigang@scut.edu.cn

Abstract

Propidium monoazide is a DNA-intercalating dye. PMA-qPCR has been reported as a novel method to detect live bacteria in complex samples. In this study, this method was used to monitor the sterilization effects of UHP, ultrasound and high PEF on Escherichia coli O157:H7. Our results showed that all three sterilization techniques are successful to kill viable E. coli O157:H7 cells under their appropriate conditions. PMA-qPCR can effectively monitor the amount of DNA released from viable E. coli O157:H7 cells, and the results from PMA-qPCR were highly consistent with those from plate counting after treatment with UHP, ultrasound and high PEF. The maximal ΔCt between PMA-qPCR and qPCR obtained in this study was 10·39 for UHP, 5·76 for ultrasound and 2·30 for high PEF. The maximal sterilization rates monitored by PMA-qPCR were 99·92% for UHP, 99·99% for ultrasound and 100% for high PEF. Thus, PMA-qPCR can be used to detect the sterilization effect on food and water supplies after treatment with UHP, ultrasound and high PEF.

Significance and Impact of the Study

The reliable detection of viable foodborne pathogenic bacteria in water and food is of great importance in our daily life. However, the traditional bacteria cultivation-based methods are time-consuming and difficult to monitor all viable bacteria because of the limitation of cultivation conditions. This study demonstrated that PMA-qPCR technique is very effective to monitor viable E. coli O157:H7 after sterilization and will help to monitor the viable bacteria in food and water.

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