Rapid Forensic Evaluation of Microbes in Biodefense Research Program Will Develop a Rapid Screening and Detection System for Multiple Bio- Threat (BT) Agents for the Department of Defense.

摘要

Global analytical approaches for forensic targets were developed and plant-fungal-viral model system was optimized for double-stranded (ds) RNA and protein signature identification. This approach developed at Indiana University of Pennsylvania (IUP) utilizes a non bio-threat plant-fungal-viral model system to develop methods and devices for rapid forensic analysis of microbes. The naturally occurring fungus Rhizoctonia solani (a common inhabitant of the soil ecosystem) was used to experiment, develop, optimize and validate bio-threat detection methods and potential device technologies outside of a controlled facility. This model system has proven to be an economical approach to knowledge acquisition that does not require an elaborate and more complex biological safety level (BSL) 3 capabilities. The fungus also harbors double stranded RNA viruses that influence the parasitic and saprobic activity of the fungus and could be potentially exploited for bioterrorism. Since the project s initiation in February 2009, significant progress has been made specifically: A rapid manufacturing protocol was developed; 13 different viral agents were characterized for their nucleic acid (milestone 1) and targetable proteins (milestone 3); universal primer for BT detection with double-stranded (ds) RNA genome has been identified and verified. The sequencing of the R. solani genome in partnership with the J. Craig Venter Institute (JCVI) is complete. So far 16 R. solani proteins have been annotated and several mutants with reduced genome complement (RGC) have been generated. Protocols for the integration of modern imaging methods into bio-defense research to elucidate mechanisms at the macro level that characterize host-microbe interactions were developed (milestone 2). Techniques used include panoramic imaging, confocal imaging, and LC/MS/MS technologies for protein signature development.