Rapid,high content screening of research colonies for polymicrobial infection

对研究菌落进行快速、高内涵的多种微生物感染筛查

• 批准号: 8455264
• 负责人: Ernest Fitch Guignon
• 金额: $ 14.44万
• 依托单位: CIENCIA, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2015-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8455264
• 关键词: Address; Affect; Agriculture; Animal Experimentation; Animals; Antibodies; Antigens; Behavior; Biological; Biological Assay; Biomedical Research; Communities; Complex; Coupled; DNA; Data; Detection; Development; Diagnosis; Diagnostic; Economic Burden; Enzyme-Linked Immunosorbent Assay; Fertility; Flow Cytometry; Future; Goals; Gold; Health; Human; Image; Immune system; Infection; Infectious Agent; Life; Magnetism; Maintenance; Masks; Measurement; Measures; Microbe; Modality; Molecular; Mus; Nature; Noise; Oligonucleotides; Phase; Population; Preparation; Printing; Productivity; Protocols documentation; RNA; Reagent; Research; Research Infrastructure; Risk; Sample Size; Sampling; Savings; Screening procedure; Sensitivity and Specificity; Signal Transduction; Staging; Surface Plasmon Resonance; Surveillance Program; System; Systems Analysis; T-Lymphocyte; Techniques; Time; United States National Institutes of Health; Viral; Work; animal colony; base; density; design; design and construction; human disease; immune function; improved; instrument; instrumentation; interest; microbial; novel; pathogen; prevent; programs; prototype; sensor; superparamagnetic beads; surface plasmon coupled emission; system architecture

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.

描述(申请人提供)：多菌感染是研究界非常关注的问题，因为它们对生产力研究小组、实验结果的可靠性以及在解开支撑我们对人类健康问题的理解的基本生物学问题方面可以取得的进展有重大影响。目前，对这些感染的诊断是使用多种技术(例如，酶联免疫吸附试验、聚合酶链式反应、流式细胞术)来完成的，这些技术费力、昂贵且耗时。因此，常规的群体监测是研究的一个重大障碍。基于表面等离子体共振(SPR)的物理现象，我们提出了一种高含量的分析方法(GCSPR和GCSPCE)。该系统中使用的传感器芯片将使用两个正交全息衍射栅，以实现两种模式下传感器芯片的同时查询。因此，我们预计将达到高灵敏度(&lt；30cfu/ml)和大动态范围(5至6个对数)。由于该系统能够使用简单的流体和廉价的芯片架构来测量生物样本中的一千多种不同的分析物，因此该系统将能够以较低的劳动力需求和极大的节省实现对生物样本的近实时测量。这一诊断系统的开发将在未来对农业动物和人类种群进行类似的诊断。我们的目标是创建一种T细胞分析系统，该系统提供高含量筛选、多参数表征能力，并采用最先进的集成样本处理，以便于使用。目前，还没有像拟议的分析系统那样能够提供广泛的功能、使用的简单性和有限的样本量要求的系统。 公共卫生相关性：研究中的多菌感染可能会影响群体健康和生产力，并可能危及实验结果的准确性。因此，旨在扩大我们对人类疾病的了解并开发更好的治疗方法和预防性治疗方法的研究面临风险。这项应用旨在开发一种高含量的检测系统，该系统能够以廉价和易于使用的格式提供快速、准确和灵敏的感染性病原体测量。使用这一系统将保护我们研究基础设施的完整性，并最终可能适应农业畜群和人类人口的诊断需求。