Automated Simultaneous Detection of Bioterrorism Agents

描述(由申请人提供)：本提案的目标是完成名为TIGER(遗传风险评估的三角识别)的传染病诊断平台的开发、工程和验证，并在疾控中心部署一个工作版本。TIGER是在DARPA赞助的一个项目中创建的，目的是开发技术来检测从环境中收集的样本中的各种生物武器制剂。TIGER采用基于电喷雾/质谱计的探测器和高通量聚合酶链式反应，并使用与雷达和卫星成像应用类似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成功地识别了关键的生物武器制剂(添加了)，如炭疽杆菌、鼠疫耶尔森菌和图拉氏弗朗西斯杆菌，而没有假阳性。TIGER还在临床样本中检测到了其他重要的医学细菌和病毒，包括化脓性链球菌和腺病毒，并对其进行了菌株分类。 虽然TIGER已经取得了关键的原则证明，但该仪器还需要进一步开发和验证NIAID/CDC优先病原体，如痘病毒科、阿雷纳病毒科、布尼亚病毒科、黄病毒科、托格维病科(尤其是VEE)、丝状病毒科以及α、β、伽马变形杆菌。为了能够在疾控中心部署，质谱学检测器必须在廉价和小样本清理的基础上开发，聚合酶链式反应机器人必须整合在专用仪器上。硬件工程完成后，将需要对所有生化试剂进行彻底的测试和验证。 我们认为，TIGER技术广泛应用的最有效途径是开发硬件、软件和生化试剂，然后将技术转移到疾控中心，我们将以协作模式与疾控中心合作开发重要应用程序。完成这些目标后，疾控中心将能够每天分析100个样本，以检测和筛选细菌和病毒类型，并在TIGER平台上执行高通量多位点序列分型(MLST)。

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.