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Biosensor Platform for Rapid Detection of E. coli in Drinking Water.

机译：用于快速检测饮用水中大肠杆菌的生物传感器平台。

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摘要

The need for rapid, specific and sensitive assays that provide a detection of bacterial indicators are important for monitoring water quality. Rapid detection using biosensor is a novel approach for microbiological testing applications. Besides, validation of rapid methods is an obstacle in adoption of such new bio-sensing technologies. In this study, the strategy developed is based on using the compound 4-methylumbelliferyl glucuronide (MUG), which is hydrolyzed rapidly by the action of E. coli beta-D-glucuronidase (GUD) enzyme to yield a fluorogenic product that can be quantified and directly related to the number of E. coli cells present in water samples. The detection time required for the biosensor response ranged from 30 to 120 minutes, depending on the number of bacteria. The specificity of the MUG based biosensor platform assay for the detection of E. coli was examined by pure cultures of non-target bacterial genera and also non-target substrates. GUD activity was found to be specific for E. coli and no such enzymatic activity was detected in other species. Moreover, the sensitivity of rapid enzymatic assays was investigated and repeatedly determined to be less than 10 E. coli cells per reaction vial concentrated from 100 mL of water samples. The applicability of the method was tested by performing fluorescence assays under pure and mixed bacterial flora in environmental samples. In addition, the procedural QA/QC for routine monitoring of drinking water samples have been validated by comparing the performance of the biosensor platform for the detection of E. coli and culture-based standard techniques such as Membrane Filtration (MF). The results of this study indicated that the fluorescence signals generated in samples using specific substrate molecules can be utilized to develop a bio-sensing platform for the detection of E. coli in drinking water. The procedural QA/QC of the biosensor will provide both industry and regulatory authorities a useful tool for near real-time monitoring of E. coli in drinking water samples. Furthermore, this system can be applied independently or in conjunction with other methods as a part of an array of biochemical assays in order to reliably detect E. coli in water.
机译：需要快速，特异性和灵敏的检测方法来检测细菌指示剂，这对于监控水质非常重要。使用生物传感器的快速检测是微生物检测应用中的一种新颖方法。此外，快速方法的验证是采用这种新的生物传感技术的障碍。在这项研究中，所开发的策略是基于使用化合物4-甲基伞形酮基葡萄糖醛酸（MUG），该化合物通过大肠杆菌β-D-葡萄糖醛酸苷酶（GUD）酶的作用而迅速水解，从而产生可以定量的荧光产物并与水样中存在的大肠杆菌细胞数量直接相关。生物传感器响应所需的检测时间范围为30到120分钟，具体取决于细菌的数量。通过基于非靶细菌属以及非靶底物的纯培养物，检测了基于MUG的基于生物传感器平台的大肠杆菌检测方法的特异性。发现GUD活性对大肠杆菌具有特异性，在其他物种中未检测到这种酶活性。此外，对快速酶法测定的敏感性进行了研究，并反复测定每个浓缩100毫升水样品的反应小瓶少于10个大肠杆菌细胞。该方法的适用性通过在环境样品中纯净的细菌和混合的细菌菌落下进行荧光分析来测试。此外，通过比较用于检测大肠杆菌的生物传感器平台和基于培养的标准技术（例如膜过滤（MF））的性能，已验证了用于饮用水样品常规监测的程序QA / QC。这项研究的结果表明，使用特定底物分子在样品中产生的荧光信号可用于开发生物传感平台，用于检测饮用水中的大肠杆菌。生物传感器的程序质量保证/质量控制将为行业和监管机构提供一个有用的工具，用于近实时监测饮用水样品中的大肠杆菌。此外，该系统可以作为一系列生化测定的一部分独立应用或与其他方法结合使用，以便可靠地检测水中的大肠杆菌。

著录项

  • 作者

    Hesari, Nikou.;

  • 作者单位

    Arizona State University.;

  • 授予单位 Arizona State University.;
  • 学科 Civil engineering.;Environmental engineering.
  • 学位 Ph.D.
  • 年度 2015
  • 页码 78 p.
  • 总页数 78
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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