Development of a universal method for diagnostic sample inactivation, extraction and enrichment of pathogens in arthropod hosts of military importance

We propose to develop an easy-to-use, safe, automated system, with related methods and consumables, to allow for the simultaneous inactivation and extraction of target molecules from pathogenic organisms, such as viruses, and rickettsia and non-rickettsial bacteria, that propagate and/or disseminate in arthropod hosts (herein referred to as Agents). The proposed System will use a combination of mechanical homogenization, alternating high hydrostatic pressure, elevated temperature, specialized processing containers, and optimized chemical reagents to inactivate said Agents and to extract the target molecules from host organisms. Following inactivation and extraction, the targets (pathogen-derived molecules such as proteins, DNA or RNA) will be fractionated and enriched for reproducible and accurate detection. We propose that the System would be equally effective in the field or in a laboratory environment. All sample processing steps will occur in individually sealed sample containers. These containers will facilitate sample homogenization, lysis, and rapid extraction of target molecules. Additionally, they will be suitable for sample collection, transportation, and storage (pre and post analysis). Such containers will help ensure strict chain-of-custody, prevent sample cross-contamination, and reduce the likelihood of exposing personnel to infectious Agents. The System will be made rugged and portable; will be capable of automating the steps of sample inactivation, fractionation, enrichment, and extraction; and will be deployable in the field environment.