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首页> 外文期刊>Biotechnology Progress >Detection of Escherichia coli with Fluorescent Labeled Phages That Have a Broad Host Range to E. coli in Sewage Water
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Detection of Escherichia coli with Fluorescent Labeled Phages That Have a Broad Host Range to E. coli in Sewage Water

机译：用荧光标记的噬菌体检测大肠杆菌，该噬菌体对污水中的大肠杆菌具有广泛的宿主范围

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Escherichia coli is used as an indicator microorganism in public health. The conventional way to detect E. coli requires several days to produce a result, because it requires incubation of cells. Therefore a rapid and sensitive detection method is needed. T4e~-/GFP phage, characterized by suppression of lysozyme and fusion of GFP (green fluorescent protein) to its SOC (small outer capsid) protein, was constructed, and it was shown to be able to detect E. coli K12 sensitively within several hours. However, because the host range of T4 phage to E. coli present in sewage water and sea water is narrow, this phage cannot be used to detect E. coli in environmental water. Two phages named IP008 and IP052, which have a broad host range to E. coli present in sewage influent, were screened from sewage influent. Mixture of these two phages produced clear plaques on 50% of E. coli screened from sewage influent. To use these phages as a tool for detection of E. coli, gfp was inserted into gene e, which encodes a lytic enzyme, and thus lytic-activity-suppressed phages were constructed (IP008e-/GFP and IP052e-/GFP). However, the fluorescent intensity of E. coli cells infected with IP008e-/GFP and IP052e-/GFP was not enough for visualization of the cell. Therefore, in addition to the insertion of gfp into gene e, fusion of GFP to SOC of IP008e-/GFP and IP052e-/GFP was conducted to produce IP008e-/ 2xGFP and IP052e-/2xGFP. E. coli cells infected with IP008e-/2xGFP and IP052e-/2xGFP showed much stronger fluorescence intensity than E. coli cells infected by IP008e-/GFP and IP052e-/GFP. It is anticipated that, using these GFP-labeled phages, a broad range of E. coli present in sewage influent water can be detected rapidly.
机译：大肠杆菌在公共卫生中被用作指示微生物。检测大肠杆菌的常规方法需要几天才能产生结果，因为它需要孵育细胞。因此，需要一种快速而灵敏的检测方法。构建了T4e〜-/ GFP噬菌体，该噬菌体的特征在于溶菌酶的抑制和GFP（绿色荧光蛋白）与其SOC（小外衣壳）蛋白的融合，并被证明能够在数个灵敏度内灵敏地检测大肠杆菌K12小时。但是，由于存在于污水和海水中的T4噬菌体对大肠杆菌的宿主范围很窄，因此该噬菌体不能用于检测环境水中的大肠杆菌。从污水入水中筛选了两个噬菌体，命名为IP008和IP052，它们对污水入水中的大肠杆菌具有广泛的宿主范围。这两种噬菌体的混合物在从污水进水中筛选出的50％的大肠杆菌上产生了清晰的噬菌斑。为了将这些噬菌体用作检测大肠杆菌的工具，将gfp插入编码裂解酶的基因e中，从而构建了抑制裂解活性的噬菌体（IP008e- / GFP和IP052e- / GFP）。但是，感染了IP008e- / GFP和IP052e- / GFP的大肠杆菌细胞的荧光强度不足以使细胞可视化。因此，除了将gfp插入基因e之外，还进行了GFP与IP008e- / GFP和IP052e- / GFP的SOC的融合以产生IP008e- / 2xGFP和IP052e- / 2xGFP。被IP008e- / 2xGFP和IP052e- / 2xGFP感染的大肠杆菌细胞显示出比被IP008e- / GFP和IP052e- / GFP感染的大肠杆菌细胞更强的荧光强度。可以预期，使用这些GFP标记的噬菌体，可以快速检测污水流入水中存在的大范围大肠杆菌。

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