Automated Simultaneous Detection of Bioterrorism Agents

自动同时检测生物恐怖分子

• 批准号: 6946857
• 负责人: David J Ecker
• 金额: $ 103.61万
• 依托单位: IONIS PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-15 至 2007-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6946857
• 关键词: DNA primers; bacterial cytopathogenic effect; bacterial genetics; biohazard detection; biomarker; biomedical automation; bioterrorism /chemical warfare; clinical research; communicable disease diagnosis; computer program /software; computer simulation; computer system design /evaluation; diagnosis design /evaluation; diagnosis quality /standard; drug resistance; electrospray ionization mass spectrometry; functional /structural genomics; high throughput technology; human tissue; polymerase chain reaction; reagent /indicator; robotics; sample collection; technology /technique development; tissue /cell preparation; virulence; virus genetics

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.

描述（由申请人提供）：该提案的目标是完成名为 TIGER（风险遗传评估三角测量识别）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 赞助的一项计划中创建的，该计划旨在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 采用基于电喷雾电离/质谱的检测器与高通量 PCR 相结合，并使用类似于雷达和卫星成像应用中使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 已成功识别出炭疽杆菌、鼠疫耶尔森氏菌和土拉弗朗西斯氏菌等关键生物武器制剂（加标），且没有出现误报。 TIGER 还对临床样本中的其他医学上重要的细菌和病毒进行了检测和菌株分型，包括化脓性链球菌和腺病毒。 虽然 TIGER 已经实现了关键的原理证明，但该仪器还需要进一步开发和验证 NIAID/CDC 优先病原体，例如痘病毒科、沙粒病毒科、布尼亚病毒科、黄病毒科、披膜病毒科（尤其是 VEE）和丝状病毒科以及 α、β、γ 病毒-变形菌。 为了能够在 CDC 进行部署，必须在廉价且小样本净化的基础上开发质谱检测器，并且必须将 PCR 机器人技术整合到专用仪器上。 硬件工程完成后，需要对所有生化试剂进行彻底的测试和验证。 我们认为，广泛利用TIGER技术的最有效途径是开发硬件、软件和生化试剂，然后将技术移交给CDC，我们将与CDC合作开发重要的应用。 完成这些目标后，CDC 将能够每天分析超过 100 个样本，以检测细菌和病毒并对其进行菌株分型，并在 TIGER 平台上执行高通量多位点序列分型 (MLST)。