New Method for Detecting Bacillus Anthracis Spores

检测炭疽芽孢杆菌孢子的新方法

• 批准号: 6561994
• 负责人: Mark S Klempner
• 金额: $ 21.71万
• 依托单位: BOSTON MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6561994
• 关键词: Bacillus anthracis; anthrax; bioterrorism /chemical warfare; microarray technology; monitoring device; spores; technology /technique development

DESCRIPTION (provided by applicant): The threat of attack on military and civilian targets with biological weapons is a growing national concern and the development of rapid, high-sensitivity, high-accuracy point detection systems for detecting and identifying bioagents in the environment and in biological specimens is a critical requirement. We will focus this effort on a new, broadly applicable method for the detection of B. anthracis spores. Currently available methods for detecting and identifying bioagents provide high sensitivity detection and highly selective identification, but the time required to chemically amplify detectable components and the size/power/cost requirement for sensor readout hardware make current methods unsuitable for point detector systems that can be placed onto any platform (including personnel or unmanned vehicles), dispersed, remotely placed, and networked. We propose to develop a revolutionary method for detecting and identifying bioagents that provides a detailed signature for a biological pathogen in real time without the use of chemical amplification. Our method relies upon measuring the affinity of bioagent outer surface components to a library of ligands immobilized onto a novel biosensor chip. Proof-of-concept of this strategy for detection, identification and discrimination of fungal spores has been demonstrated as part of a NASA funded program for the detection of microorganisms in the environment of manned space vehicles for exploratory space travel. A microarray affinity detection system that relies on both positive affinity of the agent for defined ligands as well as the lack of affinity for a separate set of ligands allows a high degree of redundancy for the specific identification of the agent as well as a powerful technology to avoid "false alarms". The sensor approach utilizes a colorimetric resonant reflectance biosensor that will allow detection hardware to be miniature, low cost, low power, and rugged. Bioagent identification is performed through automated analysis of the recognized outer surface ligands that can classify detected material (spore/bacterium/protein, alive/dead, toxic/nontoxic, pathogen family). The use of a large integrated biosensor array will provide a highly detailed bioagent signature for accurate identification with sufficient sensor redundancy to minimize false alarms. Differential signature response will be demonstrated by exposure of the microarray to vaccine strain B. anthracis spores and spores of other related and unrelated bacteria. The project will focus on developing the ligand library to include non-antibody reagents and differential recognition of bacterial spores from multiple species.

描述（由申请人提供）：用生物武器攻击军事和民用目标的威胁日益受到国家关注，快速、高灵敏度、高精度点检测系统的开发用于检测和识别环境和环境中的生物制剂。生物标本是一项关键要求。我们将把这项工作的重点放在一种新的、广泛适用的炭疽芽孢杆菌孢子检测方法上。目前可用的检测和识别生物制剂的方法提供高灵敏度检测和高选择性识别，但是化学放大可检测成分所需的时间以及传感器读出硬件的尺寸/功率/成本要求使得当前方法不适合可放置的点检测器系统任何平台（包括人员或无人驾驶车辆），分散的、远程放置的和联网的。我们建议开发一种革命性的方法来检测和识别生物制剂，无需使用化学放大即可实时提供生物病原体的详细特征。我们的方法依赖于测量生物剂外表面成分与固定在新型生物传感器芯片上的配体库的亲和力。这种检测、识别和区分真菌孢子的策略的概念验证已作为 NASA 资助的载人航天器探索性太空旅行环境中微生物检测计划的一部分进行了论证。微阵列亲和力检测系统既依赖于试剂对特定配体的正亲和力，又依赖于对一组单独的配体缺乏亲和力，从而为试剂的特异性鉴定提供了高度的冗余，并提供了强大的技术来避免“误报”。该传感器方法利用比色共振反射生物传感器，使检测硬件变得微型、低成本、低功耗且坚固耐用。生物制剂识别是通过对已识别的外表面配体进行自动分析来进行的，该外表面配体可以对检测到的材料（孢子/细菌/蛋白质、活/死、有毒/无毒、病原体家族）进行分类。使用大型集成生物传感器阵列将提供高度详细的生物制剂签名，以进行准确识别，并具有足够的传感器冗余，以最大限度地减少误报。通过将微阵列暴露于疫苗株炭疽芽孢杆菌孢子和其他相关和不相关细菌的孢子来证明差异特征反应。该项目将重点开发配体库，包括非抗体试剂和对多个物种的细菌孢子的差异识别。