A Rapid, Sensitive and Fully Automated Anthrax Test

描述（申请人提供）：生物恐怖主义和生物战是我们国家的威胁。这种威胁在2001年炭疽袭击中成为现实。这次袭击虽然规模很小，但其影响却达到了恐怖的程度。不幸的是，正如前参议员沃伦·鲁德曼所指出的那样，“未来几年我国发生恐怖袭击的可能性大于不可能性”。外交关系理事会最近的一项研究得出结论，我国仍然“没有准备好应对对美国领土的灾难性攻击，特别是涉及化学、生物或核制剂的攻击”。 在发生生物恐怖主义袭击时，最重要的第一反应之一是迅速识别生物制剂，并及时诊断受感染者。目前用于灵敏检测B的方法。炭疽是涉及聚合酶链反应（PCR）的那些。然后用培养方法和/或EIA确认微生物。然而，PCR的使用并非没有问题。PCR扩增子的可能污染需要一个专门的和高度控制的中心实验室，这反过来又需要长距离的样本运输。PCR易受抑制性杂质的影响，这一事实需要长时间的样品制备和重复检测。所有这些问题在2001年炭疽恐怖袭击期间都遇到过。 我们已经开发了一种快速（约60分钟）和敏感（少于1个细菌）的原型败血症测试使用我们的微粒为基础的扩增（MBA）技术和一类新的细菌标志物。该I期申请的具体目的是研究开发炭疽芽孢杆菌检测系统的可行性，该检测系统包括用于检测营养细胞的核酸（pXO 1和pXO 2质粒）检测和用于检测B的凝集试验。炭疽孢子两种检测方法均使用相同的MBA技术，并且具有（a）快速（60-90分钟）、（B）灵敏（核酸检测< 100 CPU，凝集检测<5000孢子）和（c）特异性（交叉反应性小）。

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).