Development of a single-use, disposable, electricity-free, nucleic acid amplification platform

Accurate diagnostic tools are essential for effective diagnosis, treatment, and tracking of individuals with infectious diseases. Diagnosis can be especially difficult in low-resource settings with limited infrastructure or unreliable access to electricity. Our previous work has demonstrated feasibility of a reusable configuration of an electricity-free, non-instrumented nucleic acid amplification platform (NINA) that complements isothermal assays for precise detection of a variety of pathogens. To maximize the potential impact of the NINA technology, we redesigned the technology in a disposable configuration to meet challenging cost, size, and ease of use requirements. This paper discusses the development of the single-use, disposable NINA platform. Key challenges included meeting repeatability, temperature stability, and precision targets. Through an iterative design process, we met these challenges using a novel design strategy including re-engineering of phase change materials (PCMs), addition of thermal enhancement additives to the PCM, and metered introduction of reactants using porous media. Manufacturing, performance, and reproducibility challenges were addressed, and the resultant design was rigorously tested in preparation for the production of a pilot batch of 150 disposable NINA devices that will be used for device and assay performance testing.