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Highly sensitive and multiplexed platforms for allergy diagnostics.

机译：用于过敏诊断的高灵敏度和多路复用平台。

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摘要

Allergy is a disorder of the immune system caused by an immune response to otherwise harmless environmental allergens. Currently 20% of the US population is allergic and 90% of pediatric patients and 60% of adult patients with asthma have allergies. These percentages have increased by 18.5% in the past decade, with predicted similar trends for the future. Here we design sensitive, multiplexed platforms to detect allergen-specific IgE using the Interferometric Reflectance Imaging Sensor (IRIS) for various clinical settings.;A microarray platform for allergy diagnosis allows for testing of specific IgE sensitivity to a multitude of allergens, while requiring only small volumes of patient blood sample. However, conventional fluorescent microarray technology is limited by i) the variation of probe immobilization, which hinders the ability to make quantitative, assertive, and statistically relevant conclusions necessary in immunodiagnostics and ii) the use of fluorophore labels, which is not suitable for some clinical applications due to the tendency of fluorophores to stick to blood particulates and require daily calibration methods. This calibrated fluorescence enhancement (CaFE) method integrates the low magnification modality of IRIS with enhanced fluorescence sensing in order to directly correlate immobilized probe (major allergens) density to allergen-specific IgE in patient serum. However, this platform only operates in processed serum samples, which is not ideal for point of care testing. Thus, a high magnification modality of IRIS was adapted as an alternative allergy diagnostic platform to automatically discriminate and size single nanoparticles bound to specific IgE in unprocessed, characterized human blood and serum samples. These features make IRIS an ideal candidate for clinical and diagnostic applications, such a POC testing.;The high magnification (nanoparticle counting) modality in conjunction with low magnification of IRIS in a combined instrument offers four significant advantages compared to existing sensing technologies: IRIS i) corrects for any variation in probe immobilization, ii) detects proteins from attomolar to nanomolar concentrations in unprocessed biological samples, iii) unambiguously discriminates nanoparticles tags on a robust and physically large sensor area, iv) detects protein targets with conjugated nanoparticle tags (~40nm diameter), which minimally affect assay kinetics compared to conventional microparticle tagging methods, and v) utilizes components that make the instrument inexpensive, robust, and portable. This platform was successfully validated on patient serum and whole blood samples with documented allergy profiles (ImmunoCAPRTM, ThermoFisher Scientific).
机译：过敏是由对其他无害的环境变应原的免疫反应引起的免疫系统疾病。目前，美国人口中有20％是过敏的，而90％的儿科患者和60％的成人哮喘患者有过敏。在过去十年中，这些百分比已增长了18.5％，并预测了未来的类似趋势。在这里，我们设计了灵敏的多路复用平台，可使用干涉反射成像传感器（IRIS）在各种临床环境中检测过敏原特异性IgE .;用于过敏诊断的微阵列平台允许测试对多种过敏原的特定IgE敏感性，而仅需少量患者血液样本。然而，常规的荧光微阵列技术受到以下限制：i）探针固定的变化，这阻碍了进行免疫诊断所必需的定量，确定和统计相关结论的能力，以及ii）使用不适合某些临床的荧光标记物由于荧光团倾向于粘附在血液颗粒上，因此需要日常校准方法。这种校准的荧光增强（CaFE）方法将IRIS的低倍放大模式与增强的荧光感应结合在一起，以便将固定的探针（主要变应原）密度直接与患者血清中的变应原特异性IgE相关联。但是，该平台只能在处理过的血清样本中运行，这对于即时检验而言并不理想。因此，IRIS的高放大倍数模式被用作替代过敏诊断平台，以自动识别和处理未加工的，特征化的人类血液和血清样品中与特定IgE结合的单个纳米颗粒。这些功能使IRIS成为临床和诊断应用（例如POC测试）的理想选择。与现有的传感技术相比，组合仪器中的高放大倍率（纳米粒子计数）模式和低放大倍率的IRIS带来了四个显着优势：IRIS i ）纠正探针固定的任何变化，ii）在未处理的生物样品中检测从摩尔浓度到纳摩尔浓度的蛋白质，iii）在坚固且物理上较大的传感器区域上明确区分纳米颗粒标签，iv）用结合的纳米颗粒标签（〜40nm）检测蛋白质靶标直径），与常规微粒标记方法相比，对分析动力学的影响最小； v）利用使仪器便宜，坚固且便携的组件。该平台已在患者血清和全血样品上成功验证，并具有已记录的过敏状况（ImmunoCAPRTM，ThermoFisher Scientific）。

著录项

  • 作者

    Monroe, Margo R.;

  • 作者单位

    Boston University.;

  • 授予单位 Boston University.;
  • 学科 Engineering Biomedical.;Nanotechnology.
  • 学位 Ph.D.
  • 年度 2013
  • 页码 175 p.
  • 总页数 175
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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