Deployable Pan-flavivirus and Pan-alphavirus Assays for Screening Pools of Medically Relevant Arthropod

CustomArray produces a field-deployable electrochemical microarray platform ideally suited for the multiplexed detection of pathogenic organisms. In this Phase II proposal, we will continue our Phase I effort to build an automated arthropod-borne virus detection system. The Phase I assay work-flow included the following three steps: (1) Extract RNA from Mosquitoes; (2) Amplify and label viral RNA loci; and (3) Detect amplification product on microarray. The results of the assay were analyzed, using software we created, to make a pathogen call. We will use this prototype detection assay as a launching pad for developing the instrumentation necessary to automate the assay and create a truly field-deployable system. In this proposal, we outline our strategy for this automation. We will (1) automate the sample prep system to extract and purify the RNA, requiring only an input of mosquitoes; (2) create an amplification system which will enable multiplexed amplification of all medically relevant arboviruses; and (3) automate the hybridization and electrochemical detection for hands free processing. We envision integrating the automated sample prep/amplification system into one disposable cartridge which will insert into a device for processing and have an output of hybridization ready material appropriate interrogation using the developed hybridizer/reader.