New Method for Detecting Bacillus Anthracis Spores

检测炭疽芽孢杆菌孢子的新方法

• 批准号: 6668682
• 负责人: Mark S Klempner
• 金额: $ 21.64万
• 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 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6668682
• 关键词: Bacillus anthracis; anthrax; biohazard detection; 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）：使用生物武器对军事和平民目标的攻击威胁是日益增长的国家关注点，并且开发了快速，高敏感性，高临界点检测系统，用于检测和识别环境中的生物代理以及生物标本中的生物是一项关键要求。我们将把这项工作集中在一种新的，广泛的适用方法上，用于检测炭疽芽孢杆菌孢子。当前可用于检测和识别生物剂的方法可提供高灵敏度检测和高度选择性的识别，但是化学放大可检测到的组件所需的时间以及传感器读数硬件的尺寸/功率/成本要求使当前方法使得对点检测器系统不适合将可以放置在任何平台上（包括人事或无人工具或无人工具或无人工具），并置于remoted，remoted，remotecles，remotecles，又有网络。我们建议开发一种革命性的方法，用于检测和识别生物代理，该生物代理在不使用化学放大的情况下实时为生物病原体提供了详细的签名。我们的方法依赖于测量生物代理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。对这种检测，鉴定和歧视真菌孢子的概念证明已被证明是NASA资助的一项旨在检测微生物在载人太空车辆环境中进行探索太空旅行环境的计划的一部分。一个微阵列亲和力检测系统依赖于代理对定义的配体的积极亲和力，以及对单独的配体的缺乏亲和力，可以高度冗余，以避免特定的代理以及强大的技术来避免“误报”。传感器方法利用比色的谐振反射率生物传感器，该反射率生物传感器将允许检测硬件是微型，低成本，低功率和坚固的。生物代理鉴定是通过对可以对检测到的材料进行分类（孢子/细菌/蛋白质，活着/死亡，有毒/无毒的病原体家族的公认的外表面配体的自动分析）进行的。大型集成生物传感器阵列的使用将提供高度详细的生物代码签名，以准确识别，并具有足够的传感器冗余，以最大程度地减少错误警报。通过将微阵列暴露于疫苗菌株B.炭疽病孢子和其他相关和无关细菌的孢子来证明差异特征反应。该项目将着重于开发配体文库，以包括非抗体试剂和对多种物种细菌孢子的差异识别。