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

We propose to develop an easy-to-use, safe, automated, field-portable high-throughput 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, high hydrostatic pressure, elevated temperature, and optimized chemical reagents to inactivate said Agents and extract the target molecules (DNA, RNA and proteins) from host organisms for subsequent separation and detection by a variety of conventional methods. We propose that the System would be equally effective in the field and in a laboratory environment. All sample processing will occur in 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.