A Rapid, Sensitive and Fully Automated Anthrax Test

快速、灵敏且全自动的炭疽测试

• 批准号: 6882976
• 负责人: XING-XIANG LI
• 金额: $ 29.44万
• 依托单位: CELLEX, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6882976
• 关键词: Bacillus anthracis; agglutination reaction; anthrax; bacterial RNA; biohazard detection; bioterrorism /chemical warfare; communicable disease diagnosis; diagnosis design /evaluation; diagnostic tests; early diagnosis; flow cytometry; luminescence; monoclonal antibody; plasmids; rapid diagnosis; spores; transfer RNA

DESCRIPTION (provided by applicant): Bioterrorism and biowarfare are a threat for our nation. Such threat became reality in the 2001 Anthrax attack. Although the attack occurred at a small scale, its impact reached terror proportion. Unfortunately, as former Senator Warren Rudman pointed out, "the likelihood of a terrorist attack in this country in the next several years is more likely than unlikely". Recent study by the Council on Foreign Relations concluded that our nation remains "unprepared to handle a catastrophic attack on American soil, particularly one involving chemical, biological or nuclear agents". In the event of a Bioterrorism attack, one of the most important first responses is rapid identification of the biological agent or agents, and timely diagnosis of those who have been infected. Current methods of choice for sensitive detection of B. anthracis are those involving polymerase chain reaction (PCR). The organism is then confirmed with culture methods and/or EIA. The use of PCR is not without problems, however. Possible contamination of PCR amplicons necessitates a specialized and highly controlled central laboratory, which in turn necessitates long distance sample shipping. The fact that PCR is susceptible to inhibitory impurities requires lengthy sample preparation and duplicate testing. All these problems had been encountered during the 2001 Anthrax terror attack. We have developed a rapid (approximately 60 minutes) and sensitive (less than 1 bacterium) prototype sepsis test using our microparticle based amplification (MBA) technology and a novel class of bacterial markers. The specific aims for this Phase I application is to study the feasibility of developing a Bacillus anthracis test system comprising a nucleic acids (pXO1 and pXO2 plasmids) test for the detection of vegetative cells and an agglutination assay for the detection of B. anthracis spores. Both assays use the same MBA technology and are (a) rapid (60-90 minutes), (b) sensitive (< 100 CPU for the nucleic acid test and <5000 spores for the agglutination assay), and (c) specific (little cross reactivity).

描述（由申请人提供）：生物恐怖主义和生物战对我们国家构成威胁。这种威胁在 2001 年炭疽袭击中成为现实。这次袭击虽然规模不大，但影响却达到了恐怖程度。不幸的是，正如前参议员沃伦·鲁德曼指出的那样，“未来几年这个国家发生恐怖袭击的可能性大于可能性”。美国外交关系委员会最近的研究得出的结论是，我们的国家仍然“没有准备好应对对美国领土的灾难性袭击，特别是涉及化学、生物或核制剂的袭击”。 在发生生物恐怖袭击时，最重要的第一反应之一是快速识别生物制剂，并及时诊断感染者。目前选择的灵敏检测炭疽芽孢杆菌的方法是涉及聚合酶链式反应 (PCR) 的方法。然后通过培养方法和/或 EIA 确认生物体。然而，PCR 的使用并非没有问题。 PCR 扩增子可能受到污染，因此需要专门且高度控制的中心实验室，而这又需要长距离样本运输。 PCR 容易受到抑制性杂质的影响，因此需要漫长的样品制备和重复测试。所有这些问题都在2001年炭疽恐怖袭击中遇到过。 我们使用基于微粒的扩增 (MBA) 技术和一类新型细菌标记物开发了一种快速（约 60 分钟）且灵敏（少于 1 个细菌）的败血症原型测试。该第一阶段应用的具体目的是研究开发炭疽芽孢杆菌测试系统的可行性，该系统包括用于检测营养细胞的核酸（pXO1和pXO2质粒）测试和用于检测炭疽芽孢杆菌孢子的凝集测定。两种检测均使用相同的 MBA 技术，并且 (a) 快速（60-90 分钟）、(b) 灵敏（核酸检测 < 100 个 CPU，凝集检测 < 5000 个孢子），以及 (c) 特异性（小）交叉反应）。