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

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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.