A Syringe-Based Biosensor to Rapidly Detect Low Levels of Escherichia coli (ECOR13) in Drinking Water Using Engineered Bacteriophages
A Syringe-Based Biosensor to Rapidly Detect Low Levels of Escherichia coli (ECOR13) in Drinking Water Using Engineered Bacteriophages
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DOI：
10.3390/s20071953
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发表时间：
2020-04-01
期刊：
影响因子：
3.9
通讯作者：
Nugen, Sam R.
中科院分区：
文献类型：
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作者：
Hinkley, Troy C.;Garing, Spencer;Nugen, Sam R.
卫生的饮用水供应是公共卫生和人类整体繁荣的无条件要求。世界卫生组织（WHO）和联合国儿童基金会（UNICEF）共同努力，已经确定了饮用水的潜在微生物和化学污染物，他们共同制定了指导方针，部分定义了大肠杆菌（E. coli）的存在。大肠杆菌）是卫生条件不佳的一个指标，对健康构成重大风险。例如E.大肠杆菌是哺乳动物胃肠道中几乎无处不在的居民，100 mL饮用水中没有可检测到的计数是世界范围内用作卫生指标的标准。目前公认的EPA方法依赖于过滤，然后在选择性培养基上生长，从样品到结果需要24-48小时。作为回应，我们开发了一种基于噬菌体的快速检测方法，其检测限能力与传统方法相当，时间不到四分之一。我们将膜过滤与使用基因工程噬菌体的选择性富集相结合，以鉴定小于20个菌落形成单位（CFU）的E。大肠杆菌在100 mL饮用水中的溶解度为5 h。膜过滤与噬菌体感染的组合产生了一种新的检测方法，该方法证明了对大量饮用水中的指示生物的快速，选择性和灵敏度检测，如世界领先的监管机构所推荐的。
A sanitized drinking water supply is an unconditional requirement for public health and the overall prosperity of humanity. Potential microbial and chemical contaminants of drinking water have been identified by a joint effort between the World Health Organization (WHO) and the United Nations Children's Fund (UNICEF), who together establish guidelines that define, in part, that the presence of Escherichia coli (E. coli) in drinking water is an indication of inadequate sanitation and a significant health risk. As E. coli is a nearly ubiquitous resident of mammalian gastrointestinal tracts, no detectable counts in 100 mL of drinking water is the standard used worldwide as an indicator of sanitation. The currently accepted EPA method relies on filtration, followed by growth on selective media, and requires 24-48 h from sample to results. In response, we developed a rapid bacteriophage-based detection assay with detection limit capabilities comparable to traditional methods in less than a quarter of the time. We coupled membrane filtration with selective enrichment using genetically engineered bacteriophages to identify less than 20 colony forming units (CFU) E. coli in 100 mL drinking water within 5 h. The combination of membrane filtration with phage infection produced a novel assay that demonstrated a rapid, selective, and sensitive detection of an indicator organism in large volumes of drinking water as recommended by the leading world regulatory authorities.