Specific single-cell isolation of Escherichia coli o157 from environmental water samples by using flow cytometry and fluorescence-activated cell sorting

Shuji Ozawa, Satoshi Okabe, Satoshi Ishii

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Contamination of food and water with pathogenic bacteria is of concern. Although culture-independent detection and quantification of pathogens is useful, isolation of pathogenic bacteria is still important when identifying the sources of pathogens. Here, we report the use of flow cytometry (FCM) and fluorescence-activated cell sorting (FACS) to specifically detect and isolate individual Escherichia coli O157:H7 cells from water samples. When present at >10 cells/mL water, target pathogen was specifically detected and isolated. The FACS-sorted E. coli O157:H7 population reflected the original population diversity, in contrast to the populations obtained by immunomagnetic separation. Relative abundance of multiple pathogenic strains is important when performing source-tracking studies; therefore, single-cell isolation with FCM-FACS can be a useful tool to obtain pathogenic bacteria for source tracking purpose.

Original languageEnglish (US)
Pages (from-to)456-461
Number of pages6
JournalFoodborne pathogens and disease
Volume13
Issue number8
DOIs
StatePublished - Aug 1 2016

Bibliographical note

Funding Information:
We thank Drs. Daisuke Sano and Ayano Kobayashi (Hokkaido University) and Dr. Nobuyuki Kijima (NARO Institute of Vegetable and Tea Science) for valuable comments, and Nora Powers (University of Minnesota) for scientific editing. This study was supported by the Grants-in-Aid for Scientific Research (23860003) from Japan Society for the Promotion of Science (JSPS), the CREST program from Japan Science and Technology Agency (JST), Kurita Water and Environment Foundation Grant, Japan, and the MnDRIVE Initiative of the University of Minnesota.

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