Anthrax Detection in Clinical Samples in 30 seconds
Click on the link below to watch a TV news clip that highlights anthrax detection in 30 seconds.
TV News Clip
In the last few years the PI has developed a very exciting assay platform technology that promises to fundamentally address two underlying physical constraints of modern assays and immunoassays, namely assay rapidity and sensitivity.
By combining the use of Metal-Enhanced Fluorescence (MEF) with low power microwave heating, we can indeed significantly increase the sensitivity of surface assays, as well as > 95 % kinetically complete the assay within a few seconds. The PI has very recently demonstrated the immense clinical significance of Microwave-Accelerated Metal-Enhanced Fluorescence (MAMEF) by developing an ultra fast and ultra bright cardiac marker assay, based on myoglobin detection. In this protein based assay, myoglobin concentrations can de detected at very low concentrations (sub clinical cut-off levels) within 20 secs, in whole blood, alleviating the need for blood separation and a 40 minute incubation period, as is clinically undertaken today. Subsequently, this technology looks to fundamentally change the way we employ current clinical assays. In this 2-year proposal we have assembled a multidisciplinary team of internationally recognized scientists to develop a less than 30 second detection platform for Anthrax. The MAMEF anthrax assay offers very high sensitivity, at detectable concentrations approaching that of Northern Blots, but within seconds, and in whole blood. The assay functions as follows: Two DNA oligonucleotide probes complimentary to different regions of the target anthrax DNA will be synthesized. One probe will be 5’- labeled with biotin, and the other with fluorescein. The DNA probes will be microwave annealed in solution to DNA targets in a bulk DNA population under reaction conditions optimized for each probe pair. The annealing mixture will then be applied to a streptavidin-coated silver surface at the same temperature, permitting capture of specific DNA:DNA hybrids through binding of the biotin moiety. After extensive washing, the amount of bound DNA will be quantified by Metal-Enhanced Fluorescence of the retained fluorescein-coupled DNA probe. After determining the optimal conditions, the assay will be tested in whole blood and the sensitivity / rapidity determined by series dilution.
Chris D. Geddes. Ph.D.
University of Maryland Biotechnology Institute