Rapid,high content screening of research colonies for polymicrobial infection

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 inboth modes. As a consequence, we anticipate reaching high levels of sensitivity (lt 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 similardiagnoses 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 researchmay 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 isat 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 integrityof our research infrastructure, and may ultimately be adaptable to diagnostic needs in agricultural herds and in the human population.