(277c) A Rapid Convection PCR-Based Diagnostic Platform

Technologies based on the polymerase chain reaction (PCR) are a gold standard for laboratory-based diagnostics. And these capabilities are enhanced through the use of multiplex reaction and target detection approaches, enabling highly specific analysis to be performed. But deployment of PCR in field-based or point-of-need settings has been limited by difficulties in adapting this instrumentation to a portable format. Here we describe the development of an analytical platform that exploits thermal convection to perform rapid PCR thermocycling in a manner amenable to operation in settings with little or no laboratory infrastructure. First, we establish a link between the PCR tube design (which governs the physics of convective flow and thermocycling) and the primer design (which regulates the PCR biochemistry). We systematically characterize this previously unexplored interplay, making it possible to identify optimal conditions that favor rapid replication of the desired target. We then extend these insights to introduce a flow-dependent correlation between replication efficiency and target GC content. Finally, we combine these insights with in-situ melting curve detection to enable robust, sensitive PCR-based analysis in a format that is readily deployable at the point-of-need.