The focus of this SBIR proposal is to demonstrate rapid, multiplexed detection of acute bacterial and viral pathogens of global importance on a single test platform. Daktaris microfluidic immunoassay platform, under development since 2009 for HIV and hepatitis C (HCV) applications, is capable of femtomolar detection of bacterial and viral antigens consistent with needs for infectious disease diagnosis with high clinical sensitivity. Moreover, by integrating sample preparation, detection, and information connectivity, seamless sample-to-answer results can be obtained by a minimally trained user starting from a single fingerstick drop of whole blood in less than 30 minutes, and transmitted worldwide for immediate action. The Daktari platform has been demonstrated and validated for HIV applications in rural fishing villages in Kasensero, Uganda and under-resourced clinics in Nairobi, Kenya. Here, we propose to extend Daktaris platform beyond fingerstick detection of HIV and HCV viral load, to create a single, multiplexed cartridge for the detection of active infection with four key pathogens: Yersinia pestis, Brucella spp., Dengue virus, and Chikungunya virus. In addition to their potential use as bioweapons, these four pathogens occur naturally, yet front-line clinicians and first responders lack useful, rapid diagnostic tools that can be deployed widely for routine outbreak surveillance in populations at risk.