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Detection of multiple pathogens by surface plasmon resonance imaging.

机译：通过表面等离子体共振成像检测多种病原体。

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摘要

The goal of this research was to determine if a surface plasmon resonance imaging device could be used to detect multiple pathogenic microorganisms that are commonly connected to food related illness. First, the dynamic range of the SPR imaging device was improved by the conversion of the device from a fixed-angle configuration to a scanning angle configuration by the addition of a computer-controlled stepper motor on the prism assembly. The enhanced dynamic range was challenged by studying the application of multiple, alternating thin polymer films of Poly(L-lysine) hydrobromide and poly(L-glutamate). The original fixed angle configuration was theoretically limited to differentiating less than 5 pairs of alternating films while the scanning angle configuration was tested to 7 pairs of alternating films without reaching the limit of dynamic range.;The scanning imaging SPR device was applied to the detection and differentiation of: E. coli O157 and two non-O157 STEC strains, E. coli O128 and E. coli O101. To functionalize the gold surface of the SPR slide with an array of antibodies, self-assembled layers of 11-mercaptoundecanoic acid, poly(ethylenimine) and ExtrAvidin were applied. A commercial biotin-conjugated anti-rabbit IgG was mated to the ExtrAvidin surface and then antibody probes were acquired directly from the three types of rabbit antiserum. The results for this methodology were disappointing, with no differences between the antibody-functionalized array spots and nonfunctionalized control spots for any of the three E. coli strains.;The methodology and goals were simplified and the SPR imaging device, restored to a fixed-angle configuration, was applied to the detection of E. coli O157, Salmonella enterica, and Listeria monocytogenes in pure cultures and in apple juice. Again using a self-assembled multilayer approach, the surface was functionalized with layers of 11-Amino-undecanethiol hydrochloride, N-succinimidyl-S-acetylthiopropionate, and streptavidin-maleimide resulting in a streptavidin surface layer. Then, purified, biotin-conjugated antibodies for each of the three target bacteria were coupled to the surface in an array configuration. This methodology was partially successful, with E. coli O157 and Salmonella enterica detected at the 108 cfu/ml level in pure culture. One of the three experimental trials resulted in evidence that Listeria monocytogenes was detected at the 108 cfu/ml level but across the three experimental trials the differences with the controls were not significant at any reasonable confidence interval. None of the target bacteria were successfully detected in apple juice.
机译：这项研究的目的是确定表面等离振子共振成像设备是否可用于检测通常与食物相关疾病相关的多种病原微生物。首先，通过在棱镜组件上添加计算机控制的步进电机，将设备从固定角度配置转换为扫描角度配置，从而改善了SPR成像设备的动态范围。通过研究聚（L-赖氨酸）氢溴酸盐和聚（L-谷氨酸）的多层交替聚合物薄膜的应用，挑战了提高的动态范围。从理论上讲，最初的固定角度配置仅限于区分少于5对交替的胶片，而扫描角度配置则针对7对交替的胶片进行了测试，而没有达到动态范围的极限。分化：大肠杆菌O157和两个非O157 STEC菌株，大肠杆菌O128和大肠杆菌O101。为了用一系列抗体功能化SPR玻片的金表面，应用了11-巯基十一烷酸，聚（乙烯亚胺）和ExtrAvidin的自组装层。将商业化的生物素偶联抗兔IgG与ExtrAvidin表面交配，然后直接从三种类型的兔抗血清中获取抗体探针。该方法学的结果令人失望，对于三种大肠杆菌菌株中的任何一种，抗体功能化的阵列斑点和非功能化的对照斑点之间都没有差异。;简化了方法和目标，将SPR成像设备还原到固定的角度配置可用于检测纯培养物和苹果汁中的大肠杆菌O157，肠炎沙门氏菌和单核细胞增生李斯特菌。再次使用自组装多层方法，将表面用11-氨基-十一烷硫醇盐酸盐，N-琥珀酰亚胺基-S-乙酰硫丙酸酯和链霉亲和素-马来酰亚胺的层官能化，得到链霉亲和素表面层。然后，将用于三种靶细菌中每一种的纯化的生物素偶联抗体以阵列形式偶联至表面。这种方法是部分成功的，在纯培养物中检测到的大肠杆菌O157和沙门氏菌为108 cfu / ml。这三项实验试验之一证明了单核细胞增生李斯特菌的检出率为108 cfu / ml，但在三项实验试验中，与对照的差异在任何合理的置信区间内均不显着。苹果汁中没有成功检测到目标细菌。

著录项

  • 作者

    Walker, Stephen P.;

  • 作者单位

    The Pennsylvania State University.;

  • 授予单位 The Pennsylvania State University.;
  • 学科 Engineering Agricultural.
  • 学位 Ph.D.
  • 年度 2010
  • 页码 109 p.
  • 总页数 109
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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