This Phase I proposal seeks to develop and demonstrate an improved first responder molecular analysis system based on a simplified methodology using aluminum oxide membrane (AOM) integrated sample preparation, amplification, and detection technology. This method has been shown to be faster, simpler, less expensive and more sensitive than existing methods. This proposal seeks to extend the applicability of this method to biodefense/ environmental contamination applications for simplified molecular analysis of relevant samples applicable to first responder settings. Phase I will broadly demonstrate the advantages of the proposed method by direct comparison to currently used sample preparation, amplification, and detection methods based on speed, sensitivity, complexity, and cost for analysis. Phase I Technical Objectives include demonstrating applicability of the AOM method for use with currently used pathogen harvesting methods from gas, liquid, and solid samples, investigation of relevant pathogen lysis conditions, investigation of RT-PCR detection of pathogen derived localized nucleic acids by the AOM method, direct comparison of the AOM method with conventional methods, and generation of Preliminary Specifications for an improved first responder molecular analysis system to be developed in a Phase II proposal. Phase II will lead to commercialization of a next generation first responder molecular analysis system.
