A Rapid, Sensitive and Fully Automated Anthrax Test

快速、灵敏且全自动的炭疽测试

• 批准号: 7035799
• 负责人: XING-XIANG LI
• 金额: $ 28.97万
• 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 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7035799
• 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贸易对我们国家是威胁。这种威胁在2001年的炭疽袭击中成为现实。尽管袭击是小规模发生的，但其影响达到了恐怖比例。不幸的是，正如前参议员沃伦·鲁德曼（Warren Rudman）指出的那样，“在未来几年内在该国发生恐怖袭击的可能性比不太可能的可能性更大”。外交关系委员会最近的研究得出的结论是，我们的国家“没有准备好应对对美国土壤的灾难性攻击，尤其是涉及化学，生物或核代理商的灾难性攻击”。 如果发生生物恐怖袭击，最重要的第一反应之一是对生物剂或药物的快速识别，以及及时诊断被感染的人。当前对炭疽芽孢杆菌敏感检测的首选方法是涉及聚合酶链反应（PCR）的方法。然后用培养方法和/或EIA确认生物体。但是，PCR的使用并非没有问题。 PCR扩增子的可能污染需要一个专业且高度控制的中央实验室，这反过来需要长途样品运输。 PCR容易受到抑制杂质的事实，需要长时间的样品制备和重复的测试。所有这些问题都是在2001年炭疽恐怖袭击期间遇到的。 我们使用基于微粒的扩增（MBA）技术和一类新型的细菌标记，开发了快速（约60分钟）和敏感（小于1细菌）败血症测试。该阶段应用的具体目的是研究开发包含核酸（PXO1和PXO2质粒）测试的炭疽芽孢杆菌测试系统的可行性，以检测营养细胞和用于检测炭疽芽孢杆菌孢子的凝集测定法。这两种测定方法都使用相同的MBA技术，并且（a）快速（60-90分钟），（b）敏感（核酸测试<100 cpu，凝集测定法<5000个孢子），以及（c）特定的（小交叉反应性）。