Phage based electrochemical detection of Escherichia coli in drinking water using affinity reporter probes
Phage based electrochemical detection of Escherichia coli in drinking water using affinity reporter probes
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DOI：
10.1039/c8an01850b
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发表时间：
2019-02-21
期刊：
影响因子：
4.2
通讯作者：
Nugen, Sam R.
中科院分区：
文献类型：
--
作者：
Wang, Danhui;Hinkley, Troy;Nugen, Sam R.
监测饮用水的粪便污染指标对于确保安全供应至关重要。建立了一种快速、灵敏检测大肠杆菌的电化学方法。大肠杆菌）。该策略是基于使用工程化噬菌体（E. coli）来分离和浓缩靶E。大肠杆菌中表达的金结合肽融合碱性磷酸酶（GBPs-ALP），使检测更加方便。融合蛋白GBPs-ALP具有酶活性和直接结合到金表面上的能力。这种结合肽介导的固定化方法提供了一种新的和简单的方法，将蛋白质固定在固体表面上，不需要翻译后修饰。E.通过使用线性扫描伏安法（LSV）电化学测量金电极上ALP的活性来确定大肠杆菌。该方法已成功应用于大肠杆菌的检测。饮用水中的大肠杆菌。我们能够检测到10（5）CFU mL（-1）的E。大肠杆菌在4小时内。预孵育9小时后，1 CFU E.检测100 mL饮用水中的大肠杆菌，总检测时间为12 h。该方法与目前的EPA方法相比具有优势，并有可能应用于检测其他食品基质中的不同细菌。
The monitoring of drinking water for indicators of fecal contamination is crucial for ensuring a safe supply. In this study, a novel electrochemical method was developed for the rapid and sensitive detection of Escherichia coli (E. coli) in drinking water. This strategy is based on the use of engineered bacteriophages (phages) to separate and concentrate target E. coli when conjugated with magnetic beads, and to facilitate the detection by expressing gold binding peptides fused alkaline phosphatase (GBPs-ALP). The fusion protein GBPs-ALP has both the enzymatic activity and the ability to directly bind onto a gold surface. This binding-peptide mediated immobilization method provided a novel and simple approach to immobilize proteins on a solid surface, requiring no post-translational modifications. The concentration of E. coli was determined by measuring the activity of the ALP on gold electrodes electrochemically using linear sweep voltammetry (LSV). This approach was successfully applied in the detection of E. coli in drinking water. We were able to detect 10(5) CFU mL(-1) of E. coli within 4 hours. After 9 hours of preincubation, 1 CFU of E. coli in 100 mL of drinking water was detected with a total assay time of 12 hours. This approach compares favorably to the current EPA method and has the potential to be applied to detect different bacteria in other food matrices.