The goal of this proposed work is to develop a rapid, automated, multi-analyte diagnostic system based on a novel microflow cytometer for screening exposure to category B pathogens. The resulting device will be portable and field deployable for rapid point-of-care and on-site diagnosis of exposure. The technology can also be used for monitoring efficacy of vaccines as they are developed. Target analytes will include, but are not limited to, Brucella, Coxiella, Salmonella, Campylobacter, E. coli, Shigella, Listeria, Cryptosporidium parvum, Giardia lamblia, and VEE. Development of the proposed diagnostic system will be two pronged, including both assay and device development. First;potential assay formats, using coded microspheres as the solid support, will be investigated, including direct detection of pathogen and indirect identification of pathogen exposure by detecting anti-pathogen antibodies. Optimized assays will be run both in buffer and spiked clinical samples. Candidate capture and tracer reagents, including antibody and non-antibody recognition molecules, will be screened with the goal of reaching physiologically relevant limits-of-detection. Magnetic nanoparticles will be used to pre-concentrate pathogens within the disposable element of the device. Second, expansion of the current microflow cytometry technology will include development and optimization of the 3-color analysis capabilities, automation of the portable system, and inclusion of pathogen sorting capability. The developed system can address of number of the areas of concern for public health including: Assays that demonstrate the highest performance for their detecting pathogens in relevant clinical samples (e.g., sensitivity, specificity, rapidity, ruggedness, ease-of-use and cost-effectiveness). Rapid sample preparation, processing, and target concentration. Improved technologies, methodologies, and reagents for simultaneous testing for multiple targets. Technologies that integrate multiple methods of parallel measurements for detection in the same platform, such as identifying nucleic acids, proteins, and other targets from agents in the same assay. Technologies targeting multiple biomarkers or signature profiles to identify human immune or other physiological responses during infection. Technology adaptable for high throughput, robotic, and automated data output and analyses at minimal cost.
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| Verbarg, Jasenka; Plath, William D; Shriver-Lake, Lisa C et al. (2013) Catch and release: integrated system for multiplexed detection of bacteria. Anal Chem 85:4944-50 |
| Golden, Joel P; Justin, Gusphyl A; Nasir, Mansoor et al. (2012) Hydrodynamic focusing--a versatile tool. Anal Bioanal Chem 402:325-35 |
| Verbarg, Jasenka; Kamgar-Parsi, Kian; Shields, Adam R et al. (2012) Spinning magnetic trap for automated microfluidic assay systems. Lab Chip 12:1793-9 |
| Smith, Joshua E; Sapsford, Kim E; Tan, Weihong et al. (2011) Optimization of antibody-conjugated magnetic nanoparticles for target preconcentration and immunoassays. Anal Biochem 410:124-32 |
| Kim, Jason S; Ligler, Frances S (2010) Utilization of microparticles in next-generation assays for microflow cytometers. Anal Bioanal Chem 398:2373-82 |
| Kim, Jason S; Taitt, Chris R; Ligler, Frances S et al. (2010) Multiplexed magnetic microsphere immunoassays for detection of pathogens in foods. Sens Instrum Food Qual Saf 4:73-81 |
| Thangawng, Abel L; Kim, Jason S; Golden, Joel P et al. (2010) A hard microflow cytometer using groove-generated sheath flow for multiplexed bead and cell assays. Anal Bioanal Chem 398:1871-81 |
| Hashemi, Nastaran; Howell Jr, Peter B; Erickson, Jeffrey S et al. (2010) Dynamic reversibility of hydrodynamic focusing for recycling sheath fluid. Lab Chip 10:1952-9 |
| Kim, Jason S; Anderson, George P; Erickson, Jeffrey S et al. (2009) Multiplexed detection of bacteria and toxins using a microflow cytometer. Anal Chem 81:5426-32 |
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