Detection, quantification, and isolation of Escherichia coli O157 by using flow cytometry and fluorescence-activated sorting

灌溉水中的病原菌可能附着在蔬菜上，并可能导致食源性疾病。为了安全的水和食物供应，应查明和消除病原体污染源。为了确定病原体的来源，必须对活病原体进行特异性检测和分离。本研究旨在建立一种利用流式细胞术和荧光激活细胞分选技术（FCM-FACS）从环境水样中特异性检测和分离病原菌的方法。我们使用荧光标记的抗O 157抗体来特异性检测E。coliO 157细胞。将检测到的细胞单独分选并在96孔板中的培养基中生长，并将它们的身份确认为E. coli O157。即使在O 157/K12细胞混合物中O 157细胞的比例低至0.01%时，我们也可以特异性地检测到O 157细胞。此外，我们可以检测到O 157细胞存在于10个细胞/ml的环境水样中。分离的细胞可以通过与已知来源的E. coliO 157数据库。

Pathogenic bacteria in irrigation water may attach to vegetables and can cause foodborne diseases. For safe water and food supply, sources of pathogen contamination should be identified and removed. To identify the sources of pathogens, specific detection and isolation of live pathogens is necessary. The objectives of this study were to develop a method to specifically detect and isolate pathogenic bacteria from environmental water samples by using flow cytometry and fluorescence-activated cell sorting (FCM-FACS). We used fluorescence-labeled anti-O157 antibodies to specifically detect E. coli O157 cells. Detected cells were individually sorted and grown in a medium in 96-well plates, and their identities were confirmed as E. coli O157. Even when the proportion of O157 cells in O157/K12 cell mixture was as low as 0.01%, we could specifically detect O157 cells. In addition, we could detect O157 cells present at 10 cells/ml in environmental water samples. Isolated cells can be used to identify their origins by comparing their characteristics to those in known-source E. coli O157 databases.