ArticlePloS one2026
ThermiQuant™ AquaStream: A portable instrument for quantitative colorimetric isothermal nucleic acid amplification reactions in paper and tube formats.
Article in PloS one, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Microfluidic paper-based analytical devices (µPADs) are an attractive format for colorimetric nucleic acid amplification tests (NAATs) because they enable low-cost, portable diagnostics in resource-limited settings. However, researchers often optimize colorimetric assays in liquid reactions in tubes before translating them to µPADs. Since both formats require separate instruments for incubation and real-time sensing, direct comparison of reactions between the two formats is difficult. To address these cross-platform limitations, we developed ThermiQuant™ AquaStream, a portable benchtop device (15 × 20 × 16 cm, ~ 5 kg; cost: USD 327) that supports seamless colorimetric loop-mediated isothermal amplification (LAMP) reactions in both µPADs and tubes under a common workflow. The system enables real-time reaction tracking (every 30 seconds) through onboard image processing, precise isothermal control (±0.5 °C) using a repurposed consumer-grade sous-vide heater, and medium-throughput (24 tubes or 42 µPADs). Testing with synthetic SARS-CoV-2 orf7ab DNA fragments demonstrated a limit of detection corresponding to a 95% probability of detection (LOD95) of 110 copies per reaction in tube (22 copies/µL) and 39 copies per reaction in µPADs (5 copies/µL), estimated using probit regression. In both formats the limit of quantification (LOQ), defined as the lowest concentration yielding a coefficient of variation (CV) of quantification time (Tq) ≤ 10%, was 250 copies/reaction resulting in a strong linear (R2 = 0.98 & R2 = 0.96 respectively for tube and µPADs) standard calibration curves. ThermiQuant™ AquaStream provides an affordable and versatile benchtop platform capable of supporting both tube- and µPAD-based colorimetric LAMP assays, serving as a proof-of-concept research tool for assay development and molecular diagnostics in One Health settings such as clinics, farms, and field environments.
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