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Colorimetric LAMP (cLAMP)-based POC nucleic acid testing is particularly promising due to its speed, sensitivity, and simple visual readout. However, there is a need for a universal, automated color detection method to improve ease of use and reduce readout subjectivity using portable devices. We proposed an AI-driven workflow for automatic detection of multiple pathogens using cLAMP in a multitube strip format. Following sample extraction, reagent rehydration, and reaction incubation, the cLAMP panel was photographed with a portable device. The algorithm performed: (1) image preprocessing to locate tubes; (2) segmentation and cleaning to extract the liquid-filled region using deep learning and Otsu's binarization, followed by pixel cleaning; (3) empty tube and invalid control checks; and (4) color detection using relative Maximum Mean Discrepancy (MMD; a nonparametric statistical distance between the hue distributions of two tube images) to identify samples. The AI workflow was evaluated on 1,636 photos from 527 tests for the differential diagnosis of three panels, for skin-tropic viruses, respiratory viral pathogens, and malaria parasites, under various lighting conditions in POC environments. The AI workflow was compared to expert-adjudicated human visual readouts as the reference standard. Rejection of invalid photos, defined as panels containing empty tubes or failed controls, achieved 98.94%, 98.29%, and 96.68% accuracy (95% CI 97.83\u221299.57%, 96.97\u221299.15%, 94.13\u221298.33%) for the three panels, respectively. Misclassified images were caused by human mistakes or extreme lighting conditions. Color identification on accepted images achieved 99.97%, 100%, and 99.69% accuracy (95% CI 99.82\u2212100% for Data set 1 and 98.27\u221299.99% for Data set 3; one-sided 95% lower bound 99.90% for Data set 2). This study demonstrates a reliable automatic color readout workflow for cLAMP-based nucleic acid detection, showing potential for integration into POC testing apps for cloud-based use.<\/jats:p>","DOI":"10.1021\/acsomega.5c12299","type":"journal-article","created":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T21:02:17Z","timestamp":1785963737000},"page":"47285-47301","source":"Crossref","is-referenced-by-count":0,"title":["AI-Driven Workflow\nfor Automated Colorimetric Isothermal\nAmplification for Robust Pathogen Detection on Portable Devices"],"prefix":"10.1021","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7376-2805","authenticated-orcid":true,"given":"Ke","family":"Xu","sequence":"first","affiliation":[{"name":"Imperial College London , , ,","place":["London, U.K., SW7 2AZ"]},{"name":"Imperial College London , , Room 7S5, Commonwealth Building, Hammersmith Hospital Campus, Du Cane Road , ,","place":["London, U.K., W12 0NN"]},{"name":"Imperial College London , , Exhibition Road , ,","place":["London, U.K., SW7 2AZ"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew L.","family":"Cavuto","sequence":"additional","affiliation":[{"name":"Imperial College London , , ,","place":["London, U.K., SW7 2AZ"]},{"name":"Imperial College London , , Room 7S5, Commonwealth Building, Hammersmith Hospital Campus, Du Cane Road , ,","place":["London, U.K., W12 0NN"]},{"name":"Imperial College London , , Exhibition Road , ,","place":["London, U.K., SW7 2AZ"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3874-8810","authenticated-orcid":true,"given":"Kenny","family":"Malpartida-Cardenas","sequence":"additional","affiliation":[{"name":"Imperial College London , , ,","place":["London, U.K., SW7 2AZ"]},{"name":"Imperial College London , , Room 7S5, Commonwealth Building, Hammersmith Hospital Campus, Du Cane Road , ,","place":["London, U.K., W12 0NN"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcus","family":"Pond","sequence":"additional","affiliation":[{"name":"Imperial College Healthcare NHS Trust , , St Mary\u2019s Hospital, Praed Street , ,","place":["London, U.K., W2 1NY"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dimbintsoa","family":"Rakotomalala Robinson","sequence":"additional","affiliation":[{"name":"MRC Unit the Gambia at the London School of Hygiene and Tropical Medicine , Atlantic Boulevard, PO Box 273 , Fajara, ,","place":["Banjul, The Gambia, 00220"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francois","family":"Kiemde","sequence":"additional","affiliation":[{"name":"Unite\u0301 de Recherche Clinique de Nanoro, Institut de Recherche En Sciences de la Sante\u0301 , Bp 218 Ouaga 11 , , ,","place":["Nanoro, Boulkiemde\u0301, Burkina Faso, 10010"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bernard","family":"Hernandez","sequence":"additional","affiliation":[{"name":"Imperial College London , , ,","place":["London, U.K., SW7 2AZ"]},{"name":"Imperial College London , , Room 7S5, Commonwealth Building, Hammersmith Hospital Campus, Du Cane Road , ,","place":["London, U.K., W12 0NN"]},{"name":"Imperial College London , , Exhibition Road , ,","place":["London, U.K., SW7 2AZ"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shiranee","family":"Sriskandan","sequence":"additional","affiliation":[{"name":"Imperial College London , , ,","place":["London, U.K., SW7 2AZ"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Umberto","family":"D\u2019Alessandro","sequence":"additional","affiliation":[{"name":"MRC Unit the Gambia at the London School of Hygiene and Tropical Medicine , Atlantic Boulevard, PO Box 273 , Fajara, ,","place":["Banjul, The Gambia, 00220"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Annette","family":"Erhart","sequence":"additional","affiliation":[{"name":"MRC Unit the Gambia at the London School of Hygiene and Tropical Medicine , Atlantic Boulevard, PO Box 273 , Fajara, ,","place":["Banjul, The Gambia, 00220"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Halidou","family":"Tinto","sequence":"additional","affiliation":[{"name":"Unite\u0301 de Recherche Clinique de Nanoro, Institut de Recherche En Sciences de la Sante\u0301 , Bp 218 Ouaga 11 , , ,","place":["Nanoro, Boulkiemde\u0301, Burkina Faso, 10010"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aubrey J.","family":"Cunnington","sequence":"additional","affiliation":[{"name":"Imperial College London , , ,","place":["London, U.K., SW7 2AZ"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alison","family":"Holmes","sequence":"additional","affiliation":[{"name":"Imperial College London , , ,","place":["London, U.K., SW7 2AZ"]},{"name":"Imperial College London , , Room 7S5, Commonwealth Building, Hammersmith Hospital Campus, Du Cane Road , ,","place":["London, U.K., W12 0NN"]},{"name":"The Fleming Initiative, Imperial College London and Imperial College Healthcare NHS Trust , St Mary\u2019s Hospital, Praed Street , ,","place":["London, U.K., W2 1NY"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pantelis","family":"Georgiou","sequence":"additional","affiliation":[{"name":"Imperial College London , , Exhibition Road , ,","place":["London, U.K., SW7 2AZ"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6689-0486","authenticated-orcid":true,"given":"Nicolas","family":"Moser","sequence":"additional","affiliation":[{"name":"Imperial College London , , Exhibition Road , ,","place":["London, U.K., SW7 2AZ"]},{"name":"The Fleming Initiative, Imperial College London and Imperial College Healthcare NHS Trust , St Mary\u2019s Hospital, Praed Street , ,","place":["London, U.K., W2 1NY"]},{"name":"Imperial College London , , ,","place":["London, U.K., SW7 2AZ"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2583-8366","authenticated-orcid":true,"given":"Jesus","family":"Rodriguez-Manzano","sequence":"additional","affiliation":[{"name":"Imperial College London , , ,","place":["London, U.K., SW7 2AZ"]},{"name":"Imperial College London , , Room 7S5, Commonwealth Building, Hammersmith Hospital Campus, Du Cane Road , ,","place":["London, U.K., W12 0NN"]},{"name":"The Fleming Initiative, Imperial College London and Imperial College Healthcare NHS Trust , St Mary\u2019s Hospital, Praed Street , ,","place":["London, U.K., W2 1NY"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,5]]},"reference":[{"key":"2026081810491659200_cit1","doi-asserted-by":"publisher","first-page":"1997","DOI":"10.1016\/S0140-6736(21)00673-5","article-title":"The Lancet\ncommission on diagnostics: transforming access to diagnostics","volume":"398","author":"Fleming","year":"2021","journal-title":"The Lancet"},{"key":"2026081810491659200_cit2","doi-asserted-by":"publisher","first-page":"828","DOI":"10.1186\/s12879-021-06528-3","article-title":"A systematic\nreview of the sensitivity and specificity of lateral flow devices\nin the detection of SARS-CoV-2","volume":"21","author":"Mistry","year":"2021","journal-title":"BMC Infect Dis."},{"key":"2026081810491659200_cit3","doi-asserted-by":"publisher","first-page":"101092","DOI":"10.1016\/j.nantod.2021.101092","article-title":"Point-of-care\ndiagnostics for infectious diseases: From methods to devices","volume":"37","author":"Wang","year":"2021","journal-title":"Nano Today"},{"key":"2026081810491659200_cit4","doi-asserted-by":"publisher","first-page":"62","DOI":"10.1007\/s10156-009-0669-9","article-title":"Loop-mediated\nisothermal amplification (LAMP): a rapid,\naccurate, and cost-effective diagnostic method for infectious diseases","volume":"15","author":"Mori","year":"2009","journal-title":"J. Infect. Chemother."},{"key":"2026081810491659200_cit5","doi-asserted-by":"publisher","first-page":"59","DOI":"10.2144\/000114253","article-title":"Visual detection\nof isothermal nucleic\nacid amplification using pH-sensitive dyes","volume":"58","author":"Tanner","year":"2015","journal-title":"BioTechniques"},{"issue":"1","key":"2026081810491659200_cit6","doi-asserted-by":"publisher","first-page":"4132","DOI":"10.1038\/s41598-022-07954-2","article-title":"A deep learning-driven low-power, accurate, and portable\nplatform for rapid detection of COVID-19 using reverse-transcription\nloop-mediated isothermal amplification","volume":"12","author":"Waheed","year":"2022","journal-title":"Sci.\nRep."},{"key":"2026081810491659200_cit7","doi-asserted-by":"publisher","first-page":"107600","DOI":"10.1016\/j.microc.2022.107600","article-title":"A new RT-LAMP-on-a-Chip\ninstrument for SARS-CoV-2 diagnostics","volume":"180","author":"Torezin Mendonc\u0327a","year":"2022","journal-title":"Microchem.\nJ."},{"key":"2026081810491659200_cit8","doi-asserted-by":"publisher","first-page":"123375","DOI":"10.1016\/j.talanta.2022.123375","article-title":"One-step colorimetric\nisothermal detection of COVID-19\nwith AI-assisted automated result analysis: A platform model for future\nemerging point-of-care RNA\/DNA disease diagnosis","volume":"249","author":"Jaroenram","year":"2022","journal-title":"Talanta"},{"key":"2026081810491659200_cit9","doi-asserted-by":"publisher","first-page":"32946","DOI":"10.1039\/D0RA05866A","article-title":"Development of a tablet PC-based\nportable device for colorimetric determination of assays including\nCOVID-19 and other pathogenic microorganisms","volume":"10","author":"Sik Yoo","year":"2020","journal-title":"RSC Adv."},{"key":"2026081810491659200_cit10","doi-asserted-by":"publisher","first-page":"972","DOI":"10.3390\/v12090972","article-title":"Artificial\nIntelligence-Assisted Loop Mediated Isothermal Amplification (AI-LAMP)\nfor rapid detection of SARS-CoV-2","volume":"12","author":"Rohaim","year":"2020","journal-title":"Viruses"},{"key":"2026081810491659200_cit11","doi-asserted-by":"publisher","first-page":"3804","DOI":"10.1039\/C9LC00797K","article-title":"A sample-to-answer,\nportable platform for rapid detection of pathogens with a smartphone\ninterface","volume":"19","author":"Ma","year":"2019","journal-title":"Lab Chip."},{"key":"2026081810491659200_cit12","doi-asserted-by":"publisher","first-page":"3036","DOI":"10.1039\/D3AN00296A","article-title":"A dual paper-based\nnucleic acid extraction method from blood in under ten minutes for\npoint-of-care diagnostics","volume":"148","author":"Malpartida-Cardenas","year":"2023","journal-title":"The Analyst"},{"key":"2026081810491659200_cit13","doi-asserted-by":"publisher","first-page":"286","DOI":"10.1016\/j.jmoldx.2018.10.004","article-title":"Rapid and sensitive\ndetection of azole-resistant Aspergillus fumigatus by tandem repeat\nloop-mediated isothermal amplification","volume":"21","author":"Yu","year":"2019","journal-title":"J. Mol.\nDiagn."},{"key":"2026081810491659200_cit14","doi-asserted-by":"publisher","first-page":"113790","DOI":"10.1016\/j.lwt.2022.113790","article-title":"A smartphone-assisted\nhigh-throughput integrated color-sensing\nplatform for the rapid detection of Campylobacter coli","volume":"167","author":"Lin","year":"2022","journal-title":"LWT"},{"key":"2026081810491659200_cit15","doi-asserted-by":"publisher","first-page":"132449","DOI":"10.1016\/j.snb.2022.132449","article-title":"Diagnosis\nof pathogen infection via a multiple-wavelength colorimetric\nsensor platform with loop-mediated isothermal amplification","volume":"370","author":"Lee","year":"2022","journal-title":"Sens. Actuators, B"},{"key":"2026081810491659200_cit16","doi-asserted-by":"publisher","first-page":"424","DOI":"10.3390\/bios12060424","article-title":"SMART-LAMP: A smartphone-operated\nhandheld device for real-time colorimetric point-of-care diagnosis\nof infectious diseases via loop-mediated isothermal amplification","volume":"12","author":"Garci\u0301a-Bernalt Diego","year":"2022","journal-title":"Biosensors"},{"key":"2026081810491659200_cit17","doi-asserted-by":"publisher","first-page":"3102","DOI":"10.1021\/acsnano.5b07338","article-title":"Reading\nout single-molecule digital RNA and DNA isothermal\namplification in nanoliter volumes with unmodified camera phones","volume":"10","author":"Rodriguez-Manzano","year":"2016","journal-title":"ACS Nano"},{"key":"2026081810491659200_cit18","doi-asserted-by":"publisher","first-page":"9009","DOI":"10.1038\/s41598-020-65899-w","article-title":"Real-time colorimetric quantitative\nmolecular detection of infectious diseases on smartphone-based diagnostic\nplatform","volume":"10","author":"Yin","year":"2020","journal-title":"Sci. Rep."},{"key":"2026081810491659200_cit19","doi-asserted-by":"publisher","first-page":"313","DOI":"10.1364\/JOSAA.29.000313","article-title":"Worldwide\nprevalence of red-green color deficiency","volume":"29","author":"Birch","year":"2012","journal-title":"J.\nOpt. Soc. Am. A"},{"key":"2026081810491659200_cit20","doi-asserted-by":"publisher","first-page":"111968","DOI":"10.1016\/j.bios.2019.111968","article-title":"A colorimetric\nloop mediated isothermal amplification (LAMP) assay based on HRP-mimicking\nmolecular beacon for the rapid detection of Vibrio parahaemolyticus","volume":"151","author":"Lee","year":"2020","journal-title":"Biosens. Bioelectron."},{"key":"2026081810491659200_cit21","doi-asserted-by":"publisher","first-page":"113330","DOI":"10.1016\/j.bios.2021.113330","article-title":"Rapid and highly sensitive one-tube colorimetric RT-LAMP\nassay for visual detection of SARS-CoV-2 RNA","volume":"187","author":"He","year":"2021","journal-title":"Biosens. Bioelectron."},{"key":"2026081810491659200_cit22","doi-asserted-by":"publisher","first-page":"307","DOI":"10.1021\/acscentsci.0c01288","article-title":"Handheld point-of-care system for rapid detection of\nSARS-CoV-2 extracted RNA in under 20 min","volume":"7","author":"Rodriguez-Manzano","year":"2021","journal-title":"ACS\nCent. Sci."},{"key":"2026081810491659200_cit23","doi-asserted-by":"publisher","first-page":"13494","DOI":"10.1021\/acs.analchem.4c01762","article-title":"Mini-program\nenabled IoT intelligent\nmolecular diagnostic device for co-detection and spatiotemporal mapping\nof infectious disease pathogens","volume":"96","author":"Wang","year":"2024","journal-title":"Anal. Chem."},{"key":"2026081810491659200_cit24","doi-asserted-by":"publisher","first-page":"120407","DOI":"10.1016\/j.talanta.2019.120407","article-title":"On-chip RT-LAMP\nand colorimetric\ndetection of the prostate cancer 3 biomarker with an integrated thermal\nand imaging box","volume":"208","author":"Wang","year":"2020","journal-title":"Talanta"},{"key":"2026081810491659200_cit25","doi-asserted-by":"publisher","first-page":"15123","DOI":"10.1038\/s41598-020-72095-3","article-title":"Quantification of\ncolorimetric isothermal amplification on the smartphone and its open-source\napp for point-of-care pathogen detection","volume":"10","author":"Nguyen","year":"2020","journal-title":"Sci.\nRep."},{"key":"2026081810491659200_cit26","doi-asserted-by":"publisher","first-page":"739983","DOI":"10.1016\/j.aquaculture.2023.739983","article-title":"AI-driven colorimetric nucleic acid\ntest for tilapia\nlake virus: A large-scale, point-of-care diagnostic model for future\nemerging diseases","volume":"577","author":"Jaroenram","year":"2023","journal-title":"Aquaculture"},{"key":"2026081810491659200_cit27","doi-asserted-by":"publisher","first-page":"745","DOI":"10.1007\/s13244-018-0645-y","article-title":"Artificial\nintelligence as a medical device in radiology: ethical and regulatory\nissues in Europe and the United States","volume":"9","author":"Pesapane","year":"2018","journal-title":"Insights\nImaging"},{"key":"2026081810491659200_cit28","doi-asserted-by":"publisher","first-page":"413","DOI":"10.1016\/j.jacr.2020.09.060","article-title":"Regulatory frameworks for development and evaluation\nof artificial intelligence-based diagnostic imaging algorithms: Summary\nand recommendations","volume":"18","author":"Larson","year":"2021","journal-title":"J. Am. Coll. Radiol."},{"key":"2026081810491659200_cit36","doi-asserted-by":"publisher","first-page":"2875","DOI":"10.1038\/s41467-025-57647-3","article-title":"Portable molecular diagnostic platform\nfor rapid point-of-care detection of mpox and other diseases","volume":"16","author":"Cavuto","year":"2025","journal-title":"Nat. Commun."},{"key":"2026081810491659200_cit30","unstructured":"ProtonDx.\n          \n          Dragonfly | point of\ncare diagnostic device. https:\/\/www.protondx.com\/humanhealth\/dragonfly. Accessed July 15, 2026."},{"key":"2026081810491659200_cit31","unstructured":"Cavuto, M. L.; Pennisi, I.; Rodriguez-Manzano, J.; Georgiou, P.\n          Lid assembly for a sample\ntube, method of using the same to collect magnetic beads, and sample\nprocessing kit. 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Chem."},{"key":"2026081810491659200_cit35","doi-asserted-by":"publisher","first-page":"4193","DOI":"10.1109\/IROS.2016.7759617","article-title":"AprilTag 2: Efficient\nand robust fiducial detection","author":"Wang","year":"2016","journal-title":"2016 IEEE\/RSJ\nInternational Conference on Intelligent Robots\nand Systems (IROS)"},{"key":"2026081810491659200_cit37","doi-asserted-by":"publisher","first-page":"8925","DOI":"10.1038\/s41467-025-64027-4","article-title":"Sensitive near point-of-care\ndetection of asymptomatic and submicroscopic Plasmodium falciparum\ninfections in African endemic countries","volume":"16","author":"Rakotomalala Robinson","year":"2025","journal-title":"Nat.\nCommun."},{"key":"2026081810491659200_cit38","unstructured":"WillionXU.\n          \n          WillionXU\/Auto_cLAMP. 2025. https:\/\/github.com\/WillionXU\/Auto_cLAMP. Accessed February\n24, 2025."},{"issue":"11","key":"2026081810491659200_cit39","doi-asserted-by":"publisher","first-page":"12993","DOI":"10.1609\/aaai.v36i11.21633","article-title":"TRACER:\nExtreme attention guided\nsalient object tracing","volume":"36","author":"Lee","year":"2022","journal-title":"Proc. AAAI Conf. Artif.\nIntell."},{"key":"2026081810491659200_cit40","unstructured":"Hugging Face.\n          \n          Carve\/tracer_b7. https:\/\/huggingface.co\/Carve\/tracer_b7. Accessed November 17, 2023."},{"key":"2026081810491659200_cit41","doi-asserted-by":"publisher","first-page":"679","DOI":"10.1109\/TPAMI.1986.4767851","article-title":"A computational\napproach to edge detection","volume":"PAMI-8","author":"Canny","year":"1986","journal-title":"IEEE Trans. Pattern\nAnal. Mach. 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