Anthrax detection in clinical samples in < 30 seconds - year 3

生物恐怖主义的性质是这样的，侵略者可能会在某个时间和地点发动袭击 旨在通过大规模伤亡来制造最大的恐怖袭击。在缺乏具体规定的情况下 智能、实时检测和诊断分析是管理 攻击的后果。在这方面，PI和他的团队最近开发了一个2 Year Marge Development Awards，一种可以检测炭疽病的超快、敏感的检测方法 基因组DNA只需几秒钟即可完成。此外，该集团还制定了高度快速的 裂解孢子样本的装置(&lt；10秒)，使样本制备和检测都能够 在1分钟内出现，分析灵敏度约为500个炭疽孢子。到目前为止，这 由PI开发的技术被认为是炭疽检测的前沿，特别是 鉴于“微波加速金属增强荧光”技术具有潜在的 可应用于多种其他生物制剂的检测，甚至可用于多种生物制剂的检测。 在这个第三年的竞争更新发展计划中，我们的目标是将我们的分析应用于临床 设置和1)用各种生物样本挑战化验以确定真实世界 敏感性，2)挑战生物基质中的分析方法，对抗国际公认的小组 3)应用新的孢子裂解技术 技术对营养体进行检测，以确定检测的敏感性和特异性，最后， 4)为了确定感染后的最早时间，新技术可以检测到孢子和 老鼠血液中的营养生物体。此外，对老鼠的研究将提供急需的数据 关于炭疽杆菌在小鼠体内的病理。

The nature of Bio-terrorism is such that an aggressor is likely to strike at a time and place calculated to induce maximum terror through mass casualties. In the absence of specific intelligence, real time detection and diagnostic assays are essential tools in managing the consequences of an attack. In this regard the PI and his team have recently developed, under a 2 year MARGE developmental award, an ultra fast and sensitive assay which can detect anthrax genomic DMA within only a few seconds. In addition, the group has also developed a highly rapid means (< 10 seconds) of lysing spore samples enabling both sample preparation and detection to occur under 1 minute, with an analytical sensitivity of about 500 anthrax spores. To date, this technology developed by the PI is thought to be at the forefront for Anthrax detection, especially given that the technology "Microwave-Accelerated Metal-Enhanced Fluorescence" has the potential to be applied to the detection of many other bio agents and even multiplexed as well. In this 3rd year competitive renewal developmental proposal, we aim to apply our assay to clinical settings and 1) challenge the assay with various biological samples to determine the real-world sensitivity, 2) challenge the assay in biological matrices against an internationally recognized panel of closely related near-neighbor strains to determine assay specificity, 3) apply the new spore lysing technology to the vegetative organism to determine the assays sensitivity and specificity and finally, 4) to determine how early after infection, the new technology can detect both spores and the vegetative organism in mouse blood. In addition, the mouse study will provide much needed data concerning the pathology of B. Anthracis in the mouse.