{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T23:35:34Z","timestamp":1786491334282,"version":"build-2736575974"},"reference-count":52,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2026,5,15]],"date-time":"2026-05-15T00:00:00Z","timestamp":1778803200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100008678","name":"Center for Produce Safety","doi-asserted-by":"publisher","award":["2023CPS11"],"award-info":[{"award-number":["2023CPS11"]}],"id":[{"id":"10.13039\/100008678","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100005826","name":"Agricultural Marketing Service","doi-asserted-by":"publisher","award":["AM22SCBPCA1133"],"award-info":[{"award-number":["AM22SCBPCA1133"]}],"id":[{"id":"10.13039\/100005826","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009168","name":"Animal and Plant Health Inspection Service","doi-asserted-by":"publisher","award":["AP22VSD&B000C022"],"award-info":[{"award-number":["AP22VSD&B000C022"]}],"id":[{"id":"10.13039\/100009168","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009168","name":"Animal and Plant Health Inspection Service","doi-asserted-by":"publisher","award":["AP23OA000000C015"],"award-info":[{"award-number":["AP23OA000000C015"]}],"id":[{"id":"10.13039\/100009168","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    Microfluidic paper-based analytical devices (\u00b5PADs) 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 \u00b5PADs. 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\u2122 AquaStream, a portable benchtop device (15\u2009\u00d7\u200920\u2009\u00d7\u200916 cm,\u2009~\u20095 kg; cost: USD 327) that supports seamless colorimetric loop-mediated isothermal amplification (LAMP) reactions in both \u00b5PADs and tubes under a common workflow. The system enables real-time reaction tracking (every 30 seconds) through onboard image processing, precise isothermal control (\u00b10.5 \u00b0C) using a repurposed consumer-grade\n                    <jats:italic>sous-vide<\/jats:italic>\n                    heater, and medium-throughput (24 tubes or 42 \u00b5PADs). Testing with synthetic SARS-CoV-2\n                    <jats:italic>orf7ab<\/jats:italic>\n                    DNA fragments demonstrated a limit of detection corresponding to a 95% probability of detection (LOD95) of 110 copies per reaction in tube (22 copies\/\u00b5L) and 39 copies per reaction in \u00b5PADs (5 copies\/\u00b5L), 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)\u2009\u2264\u200910%, was 250 copies\/reaction resulting in a strong linear (R\n                    <jats:sup>2<\/jats:sup>\n                    \u2009=\u20090.98 &amp; R\n                    <jats:sup>2<\/jats:sup>\n                    \u2009=\u20090.96 respectively for tube and \u00b5PADs) standard calibration curves. ThermiQuant\u2122 AquaStream provides an affordable and versatile benchtop platform capable of supporting both tube- and \u00b5PAD-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.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0348607","type":"journal-article","created":{"date-parts":[[2026,5,15]],"date-time":"2026-05-15T17:51:33Z","timestamp":1778867493000},"page":"e0348607","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":3,"title":["ThermiQuant\u2122 AquaStream: A portable instrument for quantitative colorimetric isothermal nucleic acid amplification reactions in paper and tube formats"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1004-5440","authenticated-orcid":true,"given":"Bibek","family":"Raut","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gopal","family":"Palla","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2620-8411","authenticated-orcid":true,"given":"Jiangshan","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Skyler","family":"Campbell","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2939-4122","authenticated-orcid":true,"given":"Virendra","family":"Kumar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kyungyeon","family":"Ra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6374-3333","authenticated-orcid":true,"given":"Mohit 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