Title: Molecular monitoring of disinfection efficacy of <i>E. coli</i> O157:H7 in bottled purified drinking water by quantitative PCR with a novel dye
Abstract: Journal of Food Processing and PreservationVolume 43, Issue 2 e13875 ORIGINAL ARTICLE Molecular monitoring of disinfection efficacy of E. coli O157:H7 in bottled purified drinking water by quantitative PCR with a novel dye Yifang Cao, Yifang Cao College of Food Science and Engineering, South China University of Technology, Guangzhou, ChinaYifang Cao and Donggen Zhou contributed equally to this work.Search for more papers by this authorDonggen Zhou, Donggen Zhou Ningbo International Travel Healthcare Center, Ningbo, ChinaYifang Cao and Donggen Zhou contributed equally to this work.Search for more papers by this authorRong Li, Rong Li Zhongshan Entry-Exit Inspection and Quarantine Bureau, Zhongshan, ChinaSearch for more papers by this authorYigang Yu, Yigang Yu College of Food Science and Engineering, South China University of Technology, Guangzhou, ChinaSearch for more papers by this authorXinglong Xiao, Corresponding Author Xinglong Xiao [email protected] orcid.org/0000-0001-5173-4459 College of Food Science and Engineering, South China University of Technology, Guangzhou, China Correspondence Xinglong Xiao, College of Food Science and Engineering, South China University of Technology, 381 Wushan Road, Tianhe Strict, Guangzhou 510640, China. Email: [email protected] Xiaofeng Li, State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, 381 Wushan Road, Tianhe Strict, Guangzhou 510640, China. Email: [email protected]Search for more papers by this authorAilian Zhou, Ailian Zhou College of Food Science and Engineering, South China University of Technology, Guangzhou, ChinaSearch for more papers by this authorDongmei Liu, Dongmei Liu College of Food Science and Engineering, South China University of Technology, Guangzhou, ChinaSearch for more papers by this authorXiaofeng Li, Corresponding Author Xiaofeng Li [email protected] State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, Guangzhou, China Correspondence Xinglong Xiao, College of Food Science and Engineering, South China University of Technology, 381 Wushan Road, Tianhe Strict, Guangzhou 510640, China. Email: [email protected] Xiaofeng Li, State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, 381 Wushan Road, Tianhe Strict, Guangzhou 510640, China. Email: [email protected]Search for more papers by this author Yifang Cao, Yifang Cao College of Food Science and Engineering, South China University of Technology, Guangzhou, ChinaYifang Cao and Donggen Zhou contributed equally to this work.Search for more papers by this authorDonggen Zhou, Donggen Zhou Ningbo International Travel Healthcare Center, Ningbo, ChinaYifang Cao and Donggen Zhou contributed equally to this work.Search for more papers by this authorRong Li, Rong Li Zhongshan Entry-Exit Inspection and Quarantine Bureau, Zhongshan, ChinaSearch for more papers by this authorYigang Yu, Yigang Yu College of Food Science and Engineering, South China University of Technology, Guangzhou, ChinaSearch for more papers by this authorXinglong Xiao, Corresponding Author Xinglong Xiao [email protected] orcid.org/0000-0001-5173-4459 College of Food Science and Engineering, South China University of Technology, Guangzhou, China Correspondence Xinglong Xiao, College of Food Science and Engineering, South China University of Technology, 381 Wushan Road, Tianhe Strict, Guangzhou 510640, China. Email: [email protected] Xiaofeng Li, State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, 381 Wushan Road, Tianhe Strict, Guangzhou 510640, China. Email: [email protected]Search for more papers by this authorAilian Zhou, Ailian Zhou College of Food Science and Engineering, South China University of Technology, Guangzhou, ChinaSearch for more papers by this authorDongmei Liu, Dongmei Liu College of Food Science and Engineering, South China University of Technology, Guangzhou, ChinaSearch for more papers by this authorXiaofeng Li, Corresponding Author Xiaofeng Li [email protected] State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, Guangzhou, China Correspondence Xinglong Xiao, College of Food Science and Engineering, South China University of Technology, 381 Wushan Road, Tianhe Strict, Guangzhou 510640, China. Email: [email protected] Xiaofeng Li, State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, 381 Wushan Road, Tianhe Strict, Guangzhou 510640, China. Email: [email protected]Search for more papers by this author First published: 09 January 2019 https://doi.org/10.1111/jfpp.13875Citations: 8 Funding information: National Key Research and Development Program of China, Grant/Award Number: 2016YFF0203204 and 2018YFD0400805; Natural Science Fund of Guangdong, Grant/Award Number: 2016A030313449; Research and Development, Grant/Award Number: 2018YFD0400805 Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Abstract A new method was developed using a novel dye thiazole orange monoazide (TOMA) combined with quantitative real-time PCR (qPCR) to detect viable Escherichia coli O157:H7 cells. Different from the commonly used PMA-qPCR assay that is based on membrane integrity, this TOMA-qPCR method is based on the concept of metabolic activity. TOMA consists of three components: a nucleic acid-intercalating moiety, a crosslinkable moiety and a linker. TOMA concentration at 50 μg/ml, 20 min incubation time, and 30 min light exposure time were suggested to use for detecting viable cells. When the inoculum concentration was 103 CFU/100 ml, TOMA-qPCR assay could completely exclude the effect of dead cells treated with heat, chlorine, or UV. Moreover, TOMA-qPCR could also be used to detect viable but nonculturable state (VBNC) cells. The result shows that TOMA-qPCR would be an alternative choice for the detection of viable cells. Practical applications To specifically detect only viable cells is of great importance in most detection of microbial diagnostics, especially for detection of foodborne pathogens such as E. coli O157:H7, which could cause health risk at low concentrations. The existing detection methods have some defects in detecting only viable cells. In this article, a novel dye was developed and combined with qPCR, to monitor sanitizing efficacy of different disinfection methods on E. coli O157:H7 at low concentrations. When detecting viable E. coli in bottled water, the established TOMA-qPCR assay completely exclude the effect of dead cells when the inoculated concentration is 103 CFU/100 ml. This method is also suitable for the detection of VBNC samples. Therefore, this method can be used to detect pathogenic bacteria with low contamination under extreme conditions such as low temperature, oligotrophic, strong radiation, and so on. CONFLICT OF INTEREST The authors have declared no conflicts of interest for this article. Citing Literature Volume43, Issue2February 2019e13875 RelatedInformation