Rapid Turn-arround Multiplex Testing: Bioweapon Agents

快速周转多重测试：生物武器制剂

• 批准号: 6696679
• 负责人: James R. Prudent
• 金额: $ 54.88万
• 依托单位: ERAGEN BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-15 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6696679
• 关键词: Bacillus anthracis; Clostridium botulinum; Crimean Congo hemorrhagic fever virus; Francisella tularensis; Yersinia pestis; biotechnology; bioterrorism /chemical warfare; clinical research; communicable disease diagnosis; diagnosis design /evaluation; gene expression; genetic screening; microorganism genetics; molecular biology information system; monitoring device; nucleic acid sequence; polymerase chain reaction; smallpox virus

DESCRIPTION (provided by applicant): The goal of this project, over Phase I and Phase II, is to develop and validate a new diagnostic platform for biowarfare detection that provides ultrafast design and implementation. Today, both the scientific and security communities believe that advances in biotechnology have increased the concern for misuse in biological weapon programs. As reports of anthrax attacks across the United States multiplied late last year, an increasing concern grew that new strains with altered genomes may appear. Therefore, new diagnostic technologies that provide quick turnaround assays to previously unknown biowarfare strains are needed. To this end, we developed a novel platform, GENE-CODE 2.0, that provides an ultraquick turn-around to real-time PCR genetic testing. GENE-CODE 2.0 employs an expanded genetic information system (AEGIS) that allows for site-specific enzymatic incorporation of reporter molecules during PCR. The platform has already been demonstrated to the commercial market for ultrasensitive quantitative anthrax detection. In Phase I we have designed and demonstrated the platform for all six CDC Category A biological terror agents. All systems were capable of detecting a single copy of a given bioweapons agent genome. In Phase II we will develop multiplexed systems to analyze multiple genetic sites within a given biowarfare agent that will allow the manufacturer to change sequence specificity in an ultrafast manner. We will combine two agent-specific detection systems with an internal positive control. Detection of multiple specific targets greatly improves confidence in the specificity of positive assay results. The internal positive control lends credence to negative assay results by confirming the proper function of assay reagents. Both of these features are especially important for bioweapons detection. Once the multiplex goal is achieved we will challenge the systems by testing them with a panel of confounding DNA samples and environmental PCR inhibitors. We will demonstrate a dehydrated reaction formulation that is stable to shipping and storage. We will also develop software tools to facilitate automation of GENE-CODE detection and quantitation of bioagents. When the other aims are met we will validate the GENE-CODE system at an external site.

描述（由申请人提供）：在第一阶段和第二阶段，该项目的目标是开发和验证一个提供超快设计和实现的生物瓦尔法检测的新诊断平台。如今，科学和安全社区都认为，生物技术的进步增加了对生物武器计划滥用的关​​注。随着去年下半年美国炭疽攻击的报道乘以倍增，越来越多的担忧越来越了解基因组可能改变的新菌株。因此，需要新的诊断技术，这些技术需要为以前未知的生物处理菌株提供快速的周转测定。为此，我们开发了一个新型的平台Gene-Code 2.0，该平台为实时PCR基因测试提供了超高的转弯。 Gene-Code 2.0采用扩展的遗传信息系统（AEGIS），该系统允许在PCR期间将位点特异性的酶促掺入记者分子。该平台已经被证明是向商业市场展示的，以进行超敏化定量炭疽检测。在第一阶段，我们设计并展示了所有六种CDC类别A生物恐怖分子的平台。所有系统都能够检测给定生物武器剂基因组的单个副本。 在第二阶段，我们将开发多路复用系统，以分析给定的生物处理剂中的多个遗传位点，该系统将允许制造商以超快方式改变序列特异性。我们将将两个特定于代理的检测系统与内部阳性对照组合结合。检测多个特定靶标可以极大地提高对阳性测定结果特异性的信心。内部阳性对照通过确认测定试剂的适当功能来为负面测定结果提供信誉。这两个特征对于生物武器检测尤其重要。一旦实现了多重目标，我们将通过使用一组混杂的DNA样品和环境PCR抑制剂来测试系统来挑战它们。我们将展示一种脱水的反应配方，该配方在运输和存储方面稳定。我们还将开发软件工具，以促进基因代码检测和生物代理定量的自动化。当满足另一个目标时，我们将在外部位点验证基因代码系统。