The goal of the proposed research is to develop a highly sensitive and specific multi-locus array diagnostic for rapid identification of Yersinia pestis, the causative agent of plague. This infectious bacterial agent is on the NIAID category A priority list as a potential biological warfare (BW) agent. In order to confirm suspicions that clinical cases may be due to purposeful aerosol dissemination of Y. pestis and, as such, may be indicative of an imminent plague epidemic, and to enable healthcare professionals to instigate immediate and effective therapeutic intervention and control measures, a rapid, sensitive, and accurate method for early detection of disease is a high priority. To meet this diagnostic challenge, the detection technology will use a polymer that exhibits intrinsic amplification of fluorescence transduction events to rapidly identify species-specific genomic, proteomic, and lipo-oligosaccharide (LOS) markers of Y. pestis. This amplifying fluorescent polymer (AFP) will be fabricated as nanoparticles, and functionalized for covalent attachment of quencher-labeled molecular or aptamer beacon probes which will trigger amplified fluorescent responses when binding of the target to the probe causes dequenching of the polymer. Nomadics is teamed with Oklahoma State University College of Veterinary Medicine (OSU CVM) researchers in an extension of a collaboration that has been ongoing for four years with a focus on detecting and identifying biological warfare agents and other pathogens of biomedical interest. This collaboration has garnered funding from the National Institute of Justice, the Department of Defense, the National Science Foundation, and other agencies, as well as non-Federal funding. Preliminary research has demonstrated many of the foundational concepts of the proposed work. For example, Nomadics and OSU CVM have implemented detection platforms using molecular and aptamer beacons based on the AFP for a number of target analytes. The proposed work further extends these demonstrated concepts, will enhance the sensitivity of the platform, address specific analytes for biodefense, and implement the technology in a diagnostic microarray. This basic approach will be immediately applicable to biodefense but will allow further extension into biomedical research, medical diagnostics, and other pathogen detection and identification applications.
