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首页> 外文学位 >An evanescent wave fiber optic biosensor for the rapid detection of pathogenic bacteria using Escherichia coli O157:H7 as a model organism for assay development.
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An evanescent wave fiber optic biosensor for the rapid detection of pathogenic bacteria using Escherichia coli O157:H7 as a model organism for assay development.

机译：e逝波光纤生物传感器，用于快速检测病原菌，使用大肠杆菌O157：H7作为模型生物进行分析开发。

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摘要

An evanescent wave fiber optic biosensor was used in an attempt to develop a system for the rapid detection of pathogenic bacteria. The principle of the biosensor is a sandwich immunoassay. Capture antibodies are immobilized on the surface of specially designed silica or polystyrene fiber-optic probes and fluorescent dye-labeled antibodies are used for generation of the specific signal. Fluorescent molecules within approximately 100–1000 run of the fiber surface are excited by the evanescent field and a portion of their emission energy recouples into the fiber, which is then collected and quantitated.; The human pathogen, Escherichia coli O157:H7 was used as a model organism for assay development. Biosensor assays were developed to detect E. coli O157:H7 in pristine buffer, one, ten, and twenty-five grain ground beef samples, and unpasteurized apple juice/cider. A sample preparation scheme for direct detection of E. coli O157:H7 in ground beef and apple/juice cider samples was developed that optimized biosensor signal response. Additionally, a rapid enrichment protocol for amplification and subsequent biosensor detection of extremely low levels (10 CFU/g) of E. coli O157:H7 in ground beef was designed. The sensitivity and specificity of the biosensor was evaluated with both silica and polystyrene waveguides in all of the sample matrices tested with spiked samples prepared using the developed sample preparation protocols.; The evanescent wave biosensor was found to be a sensitive and specific system for the detection of Escherichia coli O157:H7. Lower limits of detection ranged from 3–30 CFU/mI with the silica waveguides to 100–1000 CFU/ml/g with the polystyrene waveguides. As few as 4 CFU or 0.08 CFU/g/ml could be detected following a 6 hour enrichment. The biosensor was shown to be unique in its ability to achieve sensitive detection in relatively dirty sample homogenates. False positive results generated by non-E. coli O157:H7 organisms were minimal. Only one organism, Escherichia hermanii, showed significant cross-reactivity with the biosensor assay. Calibration curves showed the biosensor to behave as a typical sandwich immunoassay with a linear signal response when plotted on a logarithmic scale.
机译：使用van逝波光纤生物传感器来尝试开发一种用于快速检测病原细菌的系统。生物传感器的原理是夹心免疫测定。捕获抗体固定在专门设计的二氧化硅或聚苯乙烯光纤探针的表面，荧光染料标记的抗体用于产生特定信号。 van逝场在纤维表面大约100-1000范围内激发荧光分子，其一部分发射能量重新耦合到纤维中，然后对其进行收集和定量。人类病原体大肠杆菌 O157：H7被用作模型生物进行测定开发。开发了生物传感器测定法以检测 E。原始缓冲液中的O157：H7大肠杆菌，一，十和二十五粒碎牛肉样品以及未经巴氏杀菌的苹果汁/苹果酒。直接检测 E的样品制备方案。研究了牛肉和苹果/果汁苹果酒样品中的大肠杆菌O157：H7，可优化生物传感器信号响应。此外，还提供了快速富集方案，用于扩增和随后的 E极低水平（<10 CFU / g）的生物传感器检测。设计了碎牛肉中的大肠杆菌O157：H7。用二氧化硅和聚苯乙烯波导对生物样品的灵敏度和特异性进行了评估，其中所有样品样品均采用已开发的样品制备方案制备的加标样品进行测试。发现van逝波生物传感器是检测大肠杆菌 O157：H7的灵敏且特异的系统。检测下限的范围从石英波导的3–30 CFU / ml到聚苯乙烯波导的100–1000 CFU / ml / g。浓缩6小时后，可检测到低至4 CFU或0.08 CFU / g / ml。该生物传感器显示出在相对较脏的样品匀浆中实现灵敏检测的独特能力。非 E产生的假阳性结果。大肠杆菌O157：H7的微生物最少。只有一种生物体， hermanii ，在生物传感器分析中显示出明显的交叉反应性。校准曲线显示，当以对数刻度绘制时，该生物传感器表现为具有线性信号响应的典型夹心免疫测定。

著录项

  • 作者

    DeMarco, Daniel Robert.;

  • 作者单位

    University of South Florida.;

  • 授予单位 University of South Florida.;
  • 学科 Biology Microbiology.; Engineering Biomedical.
  • 学位 Ph.D.
  • 年度 2001
  • 页码 151 p.
  • 总页数 151
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 微生物学;生物医学工程;
  • 关键词

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