Advancing electricity-free molecular diagnostics at the point-of-care: Optimizing the NINA platform for a malaria LAMP assay

An increasing number of isothermal nucleic acid amplification test (NAAT) approaches are under development to liberate molecular diagnostics from the laboratory and enable efficient clinical treatment decision-making at the point-of-care (POC). Each of these approaches requires the optimization of individual reaction specifications for the most effective amplification of the target nucleic acids (NAs) including NA purity, primer/probe design and reagent composition, amplification reaction temperature window of performance, total amplification time to detect, and method of detection. As a result, significant variation exists in the critical specifications that need to be achieved. To provide better thermal management capability for NAAT POC use cases, we have improved the design of our previous NAAT enabling non-instrumented nucleic acid amplification (NINA) heater technology. Thermal modeling was used to define which specifications would have the greatest impact on overall system performance. As a result, we present an isothermal assay NINA heater platform with decreased warm-up time to assay temperature, increased thermal holdover time, and tighter adherence to the optimal assay target temperature for a malaria loop-mediated isothermal amplification assay.