Partnership for Detection of Infection by Category B Pathogens

B 类病原体感染检测合作伙伴关系

• 批准号: 8133376
• 负责人: Frances S. Ligler
• 金额: $ 75.23万
• 依托单位: U.S. NAVAL RESEARCH LABORATORY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8133376
• 关键词: Address; Antibodies; Area; Automation; Biological Assay; Biological Markers; Biosensing Techniques; Brucella; Buffers; Campylobacter; Categories; Clinical; Code; Color; Coxiella; Cryptosporidium parvum; Cytometry; Data; Detection; Development; Device or Instrument Development; Devices; Diagnosis; Diagnostic; Elements; Escherichia coli; Exposure to; Giardia lamblia; Goals; Government; Hospitals; Human; Human Resources; Immune; Infection; Infectious Agent; Laboratories; Laboratory Research; Listeria; Magnetism; Measurement; Methodology; Methods; Microspheres; Military Personnel; Monitor; Nucleic Acids; Output; Pathogen detection; Patients; Performance; Physicians' Offices; Physiological; Preparation; Process; Proteins; Public Health; Q Fever; Reagent; Research Personnel; Robotics; Running; Salmonella; Sampling; Screening procedure; Sensitivity and Specificity; Shigella; Site; Solid; Sorting - Cell Movement; System; Technology; Testing; Tracer; Training; Universities; Work; assay development; base; biodefense; cost; cost effectiveness; high throughput screening; improved; interest; microorganism; nanoparticle; novel; pathogen; pathogen exposure; point of care; point-of-care diagnostics; rapid technique; response; user-friendly; vaccine efficacy

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）： 这项工作的目标是开发一种基于新型微流式细胞仪的快速、自动化、多分析物诊断系统，用于筛查 B 类病原体的暴露情况。由此产生的设备将是便携式且可现场部署的，用于快速护理点和暴露现场诊断。该技术还可用于监测疫苗开发过程中的功效。目标分析物将包括但不限于布鲁氏菌、考克斯氏菌、沙门氏菌、弯曲杆菌、大肠杆菌、志贺氏菌、李斯特氏菌、小隐孢子虫、贾第鞭毛虫和VEE。 所提议的诊断系统的开发将是双管齐下的，包括检测和设备开发。第一的;将研究使用编码微球作为固体支持物的潜在检测形式，包括直接检测病原体和通过检测抗病原体抗体间接鉴定病原体暴露。优化的检测将在缓冲液和加标临床样品中进行。将对候选捕获和示踪试剂（包括抗体和非抗体识别分子）进行筛选，目标是达到生理相关的检测限。磁性纳米颗粒将用于将病原体预浓缩在设备的一次性元件内。其次，当前微流式细胞术技术的扩展将包括三色分析功能的开发和优化、便携式系统的自动化以及病原体分选功能。 开发的系统可以解决许多公共卫生问题，包括： 证明其在相关临床样本中检测病原体的最高性能的检测（例如灵敏度、特异性、快速性、耐用性、易用性和成本效益）。 快速样品制备、处理和目标浓度。 改进的技术、方法和试剂可同时测试多个目标。 集成多种并行测量方法以在同一平台中进行检测的技术，例如在同一测定中从试剂中识别核酸、蛋白质和其他目标。 针对多种生物标志物或特征谱的技术，用于识别感染期间的人体免疫或其他生理反应。 适用于高通量、机器人和自动化数据输出和以最低成本进行分析的技术。

项目成果

A multiwavelength microflow cytometer.

多波长微流式细胞仪。

• 发表时间: 2009
• 期刊: NRL review (1988)
• 作者: Ligler,FrancesS;Erickson,JeffreyS;Golden,JoelP;Kim,JasonS;Nasir,Mansoor;Howell,PeterJ;Thangawng,AbelL;Hilliard,LisaR;Shriver-Lake,LisaC;Anderson,GeorgeP
• 通讯作者: Anderson,GeorgeP

Hydrodynamic focusing--a versatile tool.

• DOI: 10.1007/s00216-011-5415-3
• 发表时间: 2012-01
• 期刊: ANALYTICAL AND BIOANALYTICAL CHEMISTRY
• 影响因子: 4.3
• 作者: Golden, Joel P.;Justin, Gusphyl A.;Nasir, Mansoor;Ligler, Frances S.
• 通讯作者: Ligler, Frances S.

Optimization of antibody-conjugated magnetic nanoparticles for target preconcentration and immunoassays.

• DOI: 10.1016/j.ab.2010.11.005
• 发表时间: 2011-03-01
• 期刊: Analytical biochemistry
• 影响因子: 2.9
• 作者: Smith JE;Sapsford KE;Tan W;Ligler FS
• 通讯作者: Ligler FS

Multiplexed magnetic microsphere immunoassays for detection of pathogens in foods.

• DOI: 10.1007/s11694-010-9097-x
• 发表时间: 2010-05-04
• 期刊: JOURNAL OF FOOD MEASUREMENT AND CHARACTERIZATION
• 影响因子: 3.4
• 作者: Kim, Jason S;Taitt, Chris R;Ligler, Frances S;Anderson, George P
• 通讯作者: Anderson, George P

Array Biosensor for Toxin Detection: Continued Advances.

毒素检测的阵列生物传感器：持续进展。

• DOI: 10.3390/s8128361
• 发表时间: 2008-12-15
• 期刊: Sensors (Basel, Switzerland)
• 作者: Taitt CR;Shriver-Lake LC;Ngundi MM;Ligler FS
• 通讯作者: Ligler FS