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首页> 外文学位 >Efficient separation and sensitive detection of Listeria monocytogenes using magnetic nanoparticles, microfluidics and interdigitated microelectrode based impedance immunosensor.
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Efficient separation and sensitive detection of Listeria monocytogenes using magnetic nanoparticles, microfluidics and interdigitated microelectrode based impedance immunosensor.

机译：使用磁性纳米颗粒，微流控技术和基于指状微电极的阻抗免疫传感器对单核细胞增生李斯特菌进行高效分离和灵敏检测。

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摘要

Listeria monocytogenes continues to be a major foodborne pathogen that causes food poisoning and sometimes death in immunosuppressed people and abortion in pregnant women. Nanoparticles have recently drawn attentions for use in immunomagnetic separation techniques due to their greater surface area/volume ratio and better stability against sedimentation in the absence of a magnetic field. Interdigitated microelectrodes and microfluidics make material transfer more efficient and biological/chemical interaction between the surface and solution phase much quicker. Magnetic nanoparticles (Fe 3O4) with a 30 nm diameter were functionalized with rabbit anti--L. monocytogenes antibodies via biotin--streptavidin bonds and then amalgamated with target bacterial cells to capture them during a 2 h immunoreaction. A magnetic field was applied to capture the nanoparticle-- L. monocytogenes complexes and the supernatant was removed. After a washing step, L. monocytogenes was separated from a food sample and could be ready for detection by a microfluidics and interdigitated microelectrode based impedance biosensor. Capture and separation efficiency of 75% was obtained with the magnetic nanoparticles for L. monocytogenes in phosphate buffered saline (PBS) solution. When combined with the microfluidics and interdigitated microelectrode, the lower detection limits of L. monocytogenes in pure culture and food matrices were 103 and 104 CFU/ml, respectively, which were equivalent to several bacterial cells in 34.6 nl volume of a sample injected into the microfluidic chamber. A linear correlation was found between the impedance change and target bacteria in a range of 103--10 7 CFU/ml. Equivalent circuit analysis indicated that the impedance change was mainly due to the decrease in medium resistance when L. monocytogenes cells attached to the magnetic nanoparticle--antibody conjugates in mannitol solution. The separation and detection of L. monocytogenes were not affected by presence of other foodborne bacteria. A specific, sensitive, and reproducible method using the microfluidics and interdigitated microelectrode based impedance immunosensor in couple with antibody conjugated magnetic nanoparticles was able to detect L. monocytogenes as low as 103 CFU/ml in 3 h.
机译：单核细胞增生李斯特菌仍然是一种主要的食源性病原体，可导致食物中毒，有时使免疫抑制的人死亡并导致孕妇流产。纳米颗粒由于其更大的表面积/体积比以及在没有磁场的情况下对沉积的更好的稳定性，最近已引起人们对免疫磁分离技术的关注。交叉指状的微电极和微流控技术使材料转移更有效，并且表面和溶液相之间的生物/化学相互作用更快。直径30 nm的磁性纳米颗粒（Fe 3O4）用兔抗L功能化。通过生物素-链霉亲和素键产生单核细胞增生性抗体，然后与目标细菌细胞融合以在2小时的免疫反应中捕获它们。施加磁场以捕获纳米颗粒-单核细胞增生李斯特氏菌复合物，并去除上清液。洗涤步骤后，单核细胞增生李斯特氏菌已从食品样品中分离出来，可以通过基于微流体和指状微电极的阻抗生物传感器进行检测。磁性纳米颗粒对单核细胞增生李斯特氏菌在磷酸盐缓冲盐水（PBS）溶液中的捕获和分离效率达到75％。当与微流控技术和交叉指状微电极结合使用时，纯培养物和食品基质中单核细胞增生李斯特氏菌的最低检测限分别为103和104 CFU / ml，相当于注射到样品中34.6 nl体积的几个细菌细胞。微流体室。发现阻抗变化与目标细菌之间的线性相关性在103--10 7 CFU / ml的范围内。等效电路分析表明，当甘露醇溶液中单核细胞增生李斯特氏菌细胞附着于磁性纳米颗粒-抗体结合物时，阻抗变化主要是由于中等电阻的降低引起的。单核细胞增生李斯特氏菌的分离和检测不受其他食源性细菌的影响。使用微流体技术和基于交叉指状微电极的阻抗免疫传感器结合抗体偶联的磁性纳米颗粒的特异性，灵敏性和可重现性方法能够在3小时内检测出单核细胞增生李斯特氏菌低至103 CFU / ml。

著录项

  • 作者

    Kanayeva, Damira Ashimkhanovna.;

  • 作者单位

    University of Arkansas.;

  • 授予单位 University of Arkansas.;
  • 学科 Biology Cell.;Biology Microbiology.
  • 学位 Ph.D.
  • 年度 2010
  • 页码 130 p.
  • 总页数 130
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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