Automated Simultaneous Detection of Bioterrorism Agents

描述（由申请人提供）：本提案的目标是完成被称为TIGER（遗传风险评估三角识别）的传染病诊断平台的开发、工程和验证，并在CDC部署工作版本。TIGER是在darpa赞助的项目中创建的，旨在开发从环境中收集的样本中检测各种生物武器制剂的技术。TIGER采用基于电喷雾电离/质谱的检测器，结合高通量PCR，并使用类似于雷达和卫星成像应用的最大似然数据处理器。在涉及数百个环境样本的测试中，TIGER成功地鉴定了关键的生物武器制剂（加标），如炭疽芽孢杆菌、鼠疫耶尔森菌和土拉菌弗朗西斯菌，没有出现假阳性。TIGER还在临床样本中检测并分型了其他医学上重要的细菌和病毒，包括化脓性链球菌和腺病毒。

DESCRIPTION (provided by applicant): The objective of this proposal is to complete the development, engineering, and validation of the infectious disease diagnositics platform known as TIGER (Triangulation Identification for Genetic Evaluation of Risk), and to deploy a working version at the CDC. TIGER was created in a DARPA-sponsored program to develop technology to detect a broad range of biological weapons agents in samples collected from the environment. TIGER employs an electrospray ionization/mass spectrometyr-based detector coupled with high-throughput PCR and uses a maximum-likelihood data processor similar to those used for radar and satellite imaging applications. In tests involved hundreds of environmental samples, TIGER has successfully identified, without false positives, key biological weapons agents (spiked) such as Bacillus anthracis, Yersinia pestis, and Francisella tularensis. TIGER has also detected and strain-typed other medically important bacteria and viruses including Streptococcus pyogenes and Adenovirus in clinical samples. While TIGER has achieved critical proof of principle, the instrumentation needs further development and validation for NIAID/CDC-priority pathogens such as the Poxviridae, Arenaviridae, Bunyaviridae, Flaviviridae, Togaviridae (especially VEE), and Filoviridea families and the alpha, beta, gamma-proteobacteria. To enable deployment at the CDC, the mass spectrometry detector must be developed on an inexpensive and small sample clean-up, and PCR robotics must be consolidated on a dedicated instrument. Upon completion of the hardware engineering, thorough testing and validation of all biochemical reagents will be required. We believe that the most efficient route to broad utilization of TIGER technology is to develop the hardware, software, and biochemical reagents, then transition the technology to the CDC, who we will work with in a collaborative mode to develop important applications. Upon completion of these goals, the CDC will be capable of analyzing >100 of samples per day to detect and strain-type bacteria and viruses and perform high throughput multi-locus sequence typing (MLST) on the TIGER platform.