Rapid,high content screening of research colonies for polymicrobial infection

描述（由申请人提供）：多微生物感染是研究界非常关注的问题，因为它们对研究小组的生产力、实验结果的可靠性以及在揭示支撑我们对人类健康问题理解的基本生物学问题方面可以取得的进展产生了巨大影响。这些感染的诊断目前使用多种技术（如ELISA、PCR、流式细胞术）进行，这些技术费力、昂贵且耗时。因此，常规的蜂群监测是研究的一个重大障碍。我们提出了一种基于两种模式（光栅耦合表面等离子体共振；GCSPR和光栅耦合表面等离子体耦合发射；GCSPCE）的表面等离子体共振（SPR）物理现象的高含量测定方法。本系统采用的传感器芯片将采用两个正交的全息衍射光栅，以实现两种模式下传感器芯片的同时检测。因此，我们期望达到高灵敏度（<30 CFU/ml）和大动态范围（5到6个日志）。由于该系统能够使用简单的流体和廉价的芯片架构从生物样品中测量超过一千种不同的分析物，因此该系统将能够以低劳动力需求和大幅节省的方式进行生物样品的近实时测量。该诊断系统的开发将对未来农业动物和人群的类似诊断具有相关性。目标是创建一个T细胞分析系统，提供高含量筛选，多参数表征能力，并结合最先进的集成样品处理，便于使用。目前，还没有一种系统能够像提议的分析系统那样提供广泛的功能、简单的使用和有限的样本量要求。

DESCRIPTION (provided by applicant): Polymicrobial infections are of great concern to the research community because they have a dramatic impact on the productivity research groups, on the reliability of experimental results, and on the progress that can be made in unraveling fundamental biological questions that underpin our understanding of human health issues. The diagnosis of these infections is currently done using a combination of techniques (e.g. ELISA, PCR, Flow cytometry) that are laborious, costly, and time consuming. As a consequence, routine colony surveillance represents a significant impediment to research. We propose to develop a high content assay based on the physical phenomenon of surface plasmon resonance (SPR) done in two modalities (grating coupled surface plasmon resonance; GCSPR and grating coupled surface plasmon coupled emission; GCSPCE). The sensor chip employed in this system will use two orthogonal holographic diffraction gratings to enable simultaneous sensor chip interrogation in both modes. As a consequence, we anticipate reaching high levels of sensitivity (<30 CFU/ml) and a large dynamic range (5 to 6 logs). Because the system is capable of measuring more than a thousand different analytes from a biological sample using simple fluidics and an inexpensive chip architecture, this system will be capable of near real-time measurements of biological samples with a low labor demand and at dramatic savings. Development of this diagnostics system will have future relevance in similar diagnoses in agricultural animals and in human populations. The goal is to create a T cell analysis system that offers high content screening, multi-parameter characterization capability and incorporates state-of- the-art integrated sample handling for ease-of-use. At present, there are no systems available that offer the breadth of capabilities, the simplicity of use and limited sample size requirements as the proposed analysis system. PUBLIC HEALTH RELEVANCE: Polymicrobial infections in research may affect the colony health and productivity, and may compromise the accuracy of experimental results. As a consequence, research that is designed to enlarge our understanding of human disease and to develop better therapies and preventative treatments is at risk. This application aims to develop a high content assay system that is at once able to provide rapid, accurate, and sensitive measurements of infectious agents in an inexpensive and easy to use format. Use of this system will protect the integrity of our research infrastructure, and may ultimately be adaptable to diagnostic needs in agricultural herds and in the human population.