{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T13:24:44Z","timestamp":1786973084048,"version":"build-2736575974"},"reference-count":44,"publisher":"American Chemical Society (ACS)","issue":"39","license":[{"start":{"date-parts":[[2024,9,20]],"date-time":"2024-09-20T00:00:00Z","timestamp":1726790400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100008812","name":"Defence Science and Technology Group","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/501100008812","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,10,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The Coronavirus disease 2019 (COVID-19) pandemic has supercharged innovation in the field of molecular diagnostics and led to the exploration of systems that permit the autonomous identification of airborne infectious agents. Airborne virus detection is an emerging approach for determining exposure risk, although current methods limit intervention timeliness. Here, we explore reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for one-pot detection of Severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2) (SCV2) run on membrane filters suitable for micro-air-filtration of airborne viruses. We use a design of experiments statistical framework to establish the optimal additive composition for running RT-LAMP on membrane filters. Using SCV2 liquid spike-in experiments and fluorescence detection, we show that single-pot RT-LAMP on glass fiber filters reliably detected 0.10 50% tissue culture infectious dose (TCID50) SCV2 per reaction (3600 E-gene copies) and is an order of magnitude more sensitive than conventional RT-LAMP.<\/jats:p>","DOI":"10.1021\/acsomega.4c05784","type":"journal-article","created":{"date-parts":[[2024,9,20]],"date-time":"2024-09-20T06:09:46Z","timestamp":1726812586000},"page":"40832-40840","source":"Crossref","is-referenced-by-count":3,"title":["Optimization of\nReverse Transcription Loop-Mediated\nIsothermal Amplification for In Situ Detection of SARS-CoV\u20112\nin a Micro-Air-Filtration Device Format"],"prefix":"10.1021","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-0080-8636","authenticated-orcid":true,"given":"Jacob","family":"Fry","sequence":"first","affiliation":[{"name":"The University of Melbourne , , Masson Rd , , ,","place":["Parkville, Victoria, Australia, 3010"]},{"name":"The University of Melbourne , , 792 Elizabeth Street , , ,","place":["Melbourne, Victoria, Australia, 3000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jean Y. H.","family":"Lee","sequence":"additional","affiliation":[{"name":"The University of Melbourne , , 792 Elizabeth Street , , ,","place":["Melbourne, Victoria, Australia, 3000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julie L.","family":"McAuley","sequence":"additional","affiliation":[{"name":"The University of Melbourne , , 792 Elizabeth Street , , ,","place":["Melbourne, Victoria, Australia, 3000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica L.","family":"Porter","sequence":"additional","affiliation":[{"name":"The University of Melbourne , , 792 Elizabeth Street , , ,","place":["Melbourne, Victoria, Australia, 3000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ian R.","family":"Monk","sequence":"additional","affiliation":[{"name":"The University of Melbourne , , 792 Elizabeth Street , , ,","place":["Melbourne, Victoria, Australia, 3000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samuel T.","family":"Martin","sequence":"additional","affiliation":[{"name":"The University of Melbourne , , Building 261\/203 Bouverie St , , ,","place":["Carlton, Victoria, Australia, 3053"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5382-9718","authenticated-orcid":true,"given":"David J.","family":"Collins","sequence":"additional","affiliation":[{"name":"The University of Melbourne , , Building 261\/203 Bouverie St , , ,","place":["Carlton, Victoria, Australia, 3053"]},{"name":"The University of Melbourne , , Chemical Engineering 2 Building 167 , , ,","place":["Parkville, Victoria, Australia, 3010"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gregory J.","family":"Barbante","sequence":"additional","affiliation":[{"name":"Australian Department of Defence , , 506 Lorimer Street , , ,","place":["Fishermans Bend, Victoria, Australia, 3207"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicholas J.","family":"Fitzgerald","sequence":"additional","affiliation":[{"name":"Australian Department of Defence , , 506 Lorimer Street , , ,","place":["Fishermans Bend, Victoria, Australia, 3207"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0150-123X","authenticated-orcid":true,"given":"Timothy P.","family":"Stinear","sequence":"additional","affiliation":[{"name":"The University of Melbourne , , 792 Elizabeth Street , , ,","place":["Melbourne, Victoria, Australia, 3000"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2024,9,20]]},"reference":[{"key":"2026081708293629400_cit1","doi-asserted-by":"publisher","DOI":"10.1016\/j.talanta.2022.123409","article-title":"Review of COVID-19 Testing and Diagnostic\nMethods","volume":"244","author":"Filchakova","year":"2022","journal-title":"Talanta"},{"issue":"12","key":"2026081708293629400_cit2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/28.12.e63","article-title":"Loop-Mediated\nIsothermal Amplification\nof DNA","volume":"28","author":"Notomi","year":"2000","journal-title":"Nucleic Acids Res."},{"issue":"3","key":"2026081708293629400_cit3","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1006\/mcpr.2002.0415","article-title":"Accelerated Reaction\nby Loop-Mediated\nIsothermal Amplification Using Loop Primers","volume":"16","author":"Nagamine","year":"2002","journal-title":"Mol. Cell. Probes"},{"key":"2026081708293629400_cit4","doi-asserted-by":"publisher","first-page":"102","DOI":"10.1016\/j.jviromet.2017.08.013","article-title":"Defining the Relative Performance of Isothermal Assays\nThat Can Be Used for Rapid and Sensitive Detection of Foot-and-Mouth\nDisease Virus","volume":"249","author":"Howson","year":"2017","journal-title":"J. Virol. Methods"},{"issue":"1","key":"2026081708293629400_cit5","doi-asserted-by":"publisher","first-page":"257","DOI":"10.1128\/JCM.42.1.257-263.2004","article-title":"Real-Time\nReverse Transcription Loop-Mediated Isothermal Amplification for Rapid\nDetection of West Nile Virus","volume":"42","author":"Parida","year":"2004","journal-title":"J. Clin. Microbiol."},{"issue":"9","key":"2026081708293629400_cit6","doi-asserted-by":"publisher","first-page":"1713","DOI":"10.1007\/s00705-003-0134-5","article-title":"Detection of Japanese Yam Mosaic\nVirus by RT-LAMP","volume":"148","author":"Fukuta","year":"2003","journal-title":"Arch. Virol."},{"issue":"11","key":"2026081708293629400_cit7","doi-asserted-by":"publisher","DOI":"10.1016\/j.sjbs.2022.103465","article-title":"SARS-CoV-2\nDetection Methods: A Comprehensive Review","volume":"29","author":"Alhamid","year":"2022","journal-title":"Saudi\nJ. Biol. Sci."},{"issue":"5","key":"2026081708293629400_cit8","doi-asserted-by":"publisher","first-page":"877","DOI":"10.1038\/nprot.2008.57","article-title":"Loop-Mediated\nIsothermal\nAmplification (LAMP) of Gene Sequences and Simple Visual Detection\nof Products","volume":"3","author":"Tomita","year":"2008","journal-title":"Nat. Protoc."},{"issue":"1","key":"2026081708293629400_cit9","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-021-94652-0","article-title":"An Ultra-Portable,\nSelf-Contained Point-of-Care Nucleic Acid Amplification\nTest for Diagnosis of Active COVID-19 Infection","volume":"11","author":"Deng","year":"2021","journal-title":"Sci. Rep."},{"issue":"10","key":"2026081708293629400_cit10","doi-asserted-by":"publisher","first-page":"4982","DOI":"10.1021\/acsbiomaterials.1c01105","article-title":"Low-Cost, Open-Source\nDevice for\nHigh-Performance Fluorescence Detection of Isothermal Nucleic Acid\nAmplification Reactions","volume":"7","author":"Buultjens","year":"2021","journal-title":"ACS Biomater. Sci.\nEng."},{"key":"2026081708293629400_cit11","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-30797-9","article-title":"Simpler, Faster, and Sensitive Zika\nVirus Assay Using Smartphone Detection of Loop-Mediated Isothermal\nAmplification on Paper Microfluidic Chips","volume":"8","author":"Kaarj","year":"2018","journal-title":"Sci.\nRep."},{"key":"2026081708293629400_cit12","doi-asserted-by":"publisher","first-page":"774","DOI":"10.1016\/j.bios.2015.10.032","article-title":"Fabrication\nTechniques for Microfluidic Paper-Based\nAnalytical Devices and Their Applications for Biological Testing:\nA Review","volume":"77","author":"Xia","year":"2016","journal-title":"Biosens. Bioelectron."},{"issue":"6","key":"2026081708293629400_cit13","doi-asserted-by":"publisher","first-page":"8319","DOI":"10.3390\/s120608319","article-title":"Diagnostic Devices\nfor Isothermal Nucleic Acid Amplification","volume":"12","author":"Chang","year":"2012","journal-title":"Sensors"},{"issue":"3","key":"2026081708293629400_cit14","doi-asserted-by":"publisher","first-page":"611","DOI":"10.1039\/C5LC01388G","article-title":"An Integrated Paper-Based Sample-to-Answer Biosensor for Nucleic\nAcid Testing at the Point of Care","volume":"16","author":"Choi","year":"2016","journal-title":"Lab Chip"},{"key":"2026081708293629400_cit15","doi-asserted-by":"publisher","DOI":"10.1016\/j.aca.2021.338220","article-title":"Paper-Based Loop-Mediated\nIsothermal Amplification and Lateral Flow\n(LAMP-LF) Assay for Identification of Tissues of Cattle Origin","volume":"1150","author":"Jawla","year":"2021","journal-title":"Anal. Chim. Acta"},{"issue":"2","key":"2026081708293629400_cit16","doi-asserted-by":"publisher","first-page":"403","DOI":"10.1021\/acssensors.7b00825","article-title":"Rapid Veterinary Diagnosis of Bovine\nReproductive Infectious\nDiseases from Semen Using Paper-Origami DNA Microfluidics","volume":"3","author":"Yang","year":"2018","journal-title":"ACS Sens."},{"key":"2026081708293629400_cit17","doi-asserted-by":"publisher","DOI":"10.1016\/j.aca.2021.338542","article-title":"Integration\nof Sample\nPreparation with RNA-Amplification in a Hand-Held Device for Airborne\nVirus Detection","volume":"1165","author":"Jiang","year":"2021","journal-title":"Anal. Chim. Acta"},{"issue":"11","key":"2026081708293629400_cit18","doi-asserted-by":"publisher","first-page":"4834","DOI":"10.1073\/pnas.1812296116","article-title":"Paper-Based Microfluidics for DNA\nDiagnostics of Malaria in Low Resource Underserved Rural Communities","volume":"116","author":"Reboud","year":"2019","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"issue":"8","key":"2026081708293629400_cit19","doi-asserted-by":"publisher","first-page":"2220","DOI":"10.7150\/thno.18675","article-title":"A Paper-Based Device\nfor Performing Loop-Mediated Isothermal Amplification with Real-Time\nSimultaneous Detection of Multiple DNA Targets","volume":"7","author":"Seok","year":"2017","journal-title":"Theranostics"},{"key":"2026081708293629400_cit20","doi-asserted-by":"publisher","first-page":"1169","DOI":"10.1099\/jmm.0.001238","article-title":"Validation of a\nSingle-Step, Single-Tube Reverse Transcription Loop-Mediated Isothermal\nAmplification Assay for Rapid Detection of SARS-CoV-2 RNA","volume":"69","author":"Lee","year":"2020","journal-title":"J. Med. Microbiol."},{"issue":"1","key":"2026081708293629400_cit21","doi-asserted-by":"publisher","first-page":"117","DOI":"10.1016\/j.jinf.2020.10.039","article-title":"A Highly\nEffective Reverse-Transcription Loop-Mediated Isothermal Amplification\n(RT-LAMP) Assay for the Rapid Detection of SARS-CoV-2 Infection","volume":"82","author":"Fowler","year":"2021","journal-title":"J. Infect."},{"issue":"8","key":"2026081708293629400_cit22","doi-asserted-by":"publisher","first-page":"2826","DOI":"10.3390\/ijms21082826","article-title":"A Novel Reverse\nTranscription Loop-Mediated Isothermal Amplification Method for Rapid\nDetection of SARS-CoV-2","volume":"21","author":"Lu","year":"2020","journal-title":"Int. J. Mol. Sci."},{"issue":"1","key":"2026081708293629400_cit23","doi-asserted-by":"publisher","first-page":"998","DOI":"10.1080\/22221751.2020.1756698","article-title":"Development of a\nReverse Transcription-Loop-Mediated Isothermal Amplification as a\nRapid Early-Detection Method for Novel SARS-CoV-2","volume":"9","author":"Baek","year":"2020","journal-title":"Emerging Microbes Infect."},{"issue":"1","key":"2026081708293629400_cit24","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0243712","article-title":"Performing Point-of-Care Molecular\nTesting for SARS-CoV-2\nwith RNA Extraction and Isothermal Amplification","volume":"16","author":"Garneret","year":"2021","journal-title":"PLoS One"},{"issue":"4","key":"2026081708293629400_cit25","doi-asserted-by":"publisher","first-page":"364","DOI":"10.1007\/s12560-022-09522-3","article-title":"Integration\nof RT-LAMP and Microfluidic Technology for Detection of SARS-CoV-2\nin Wastewater as an Advanced Point-of-Care Platform","volume":"14","author":"Donia","year":"2022","journal-title":"Food Environ. Virol."},{"issue":"17","key":"2026081708293629400_cit26","doi-asserted-by":"publisher","first-page":"3157","DOI":"10.1039\/D2LC00242F","article-title":"Automated Sample-to-Answer\nCentrifugal Microfluidic System for Rapid\nMolecular Diagnostics of SARS-CoV-2","volume":"22","author":"Malic","year":"2022","journal-title":"Lab Chip"},{"issue":"37","key":"2026081708293629400_cit27","doi-asserted-by":"publisher","first-page":"22727","DOI":"10.1073\/pnas.2014739117","article-title":"Rapid Isothermal\nAmplification and\nPortable Detection System for SARS-CoV-2","volume":"117","author":"Ganguli","year":"2020","journal-title":"Proc.\nNatl. Acad. Sci. U.S.A."},{"issue":"4","key":"2026081708293629400_cit28","doi-asserted-by":"publisher","first-page":"e132","DOI":"10.1309\/LMMHGYKY7LIUEEQ6","article-title":"Barriers to\nthe Introduction of New Medical Diagnostic Tests","volume":"44","author":"Ivanov","year":"2013","journal-title":"Lab. Med."},{"key":"2026081708293629400_cit29","doi-asserted-by":"publisher","DOI":"10.1016\/j.scitotenv.2021.150060","article-title":"Wastewater Surveillance to Infer COVID-19 Transmission:\nA Systematic\nReview","volume":"804","author":"Shah","year":"2022","journal-title":"Sci. Total Environ."},{"key":"2026081708293629400_cit30","doi-asserted-by":"publisher","DOI":"10.1016\/j.envint.2020.106039","article-title":"Aerosol Transmission of SARS-CoV-2? Evidence, Prevention and Control","volume":"144","author":"Tang","year":"2020","journal-title":"Environ. Int."},{"key":"2026081708293629400_cit31","doi-asserted-by":"publisher","DOI":"10.1016\/j.envint.2020.105730","article-title":"Airborne Transmission of SARS-CoV-2:\nThe World Should Face the Reality","volume":"139","author":"Morawska","year":"2020","journal-title":"Environ.\nInt."},{"issue":"6","key":"2026081708293629400_cit32","doi-asserted-by":"publisher","first-page":"1167","DOI":"10.4209\/aaqr.2020.05.0202","article-title":"Collection of SARS-CoV-2\nVirus from the Air of a Clinic Within a University Student Health\nCare Center and Analyses of the Viral Genomic Sequence","volume":"20","author":"Lednicky","year":"2020","journal-title":"Aerosol Air Qual. Res."},{"issue":"9","key":"2026081708293629400_cit33","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0258151","article-title":"Aerosol SARS-CoV-2\nin Hospitals and\nLong-Term Care Homes during the COVID-19 Pandemic","volume":"16","author":"Mallach","year":"2021","journal-title":"PLoS One"},{"issue":"1","key":"2026081708293629400_cit34","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-16670-2","article-title":"Singapore\n2019 Novel Coronavirus Outbreak Research Team. Detection of Air and\nSurface Contamination by SARS-CoV-2 in Hospital Rooms of Infected\nPatients","volume":"11","author":"Chia","year":"2020","journal-title":"Nat. Commun."},{"issue":"2","key":"2026081708293629400_cit35","doi-asserted-by":"publisher","first-page":"144","DOI":"10.1038\/s41587-021-00816-8","article-title":"Research Priorities\nfor COVID-19 Sensor Technology","volume":"39","author":"Tong","year":"2021","journal-title":"Nat. Biotechnol."},{"issue":"10","key":"2026081708293629400_cit36","doi-asserted-by":"publisher","first-page":"459","DOI":"10.5694\/mja2.50569","article-title":"Isolation and Rapid\nSharing of the 2019 Novel Coronavirus (SARS-CoV-2) from the First\nPatient Diagnosed with COVID-19 in Australia","volume":"212","author":"Caly","year":"2020","journal-title":"Med. J. Aust."},{"issue":"3","key":"2026081708293629400_cit37","doi-asserted-by":"publisher","first-page":"493","DOI":"10.1093\/oxfordjournals.aje.a118408","article-title":"A Simple method of\nestimating fifty per cent endpoints12","volume":"27","author":"Reed","year":"1938","journal-title":"Am. J.\nEpidemiol."},{"issue":"9","key":"2026081708293629400_cit38","doi-asserted-by":"publisher","first-page":"4669","DOI":"10.1021\/acsbiomaterials.1c00775","article-title":"Accessible Platform\nfor High-Throughput COVID-19 Molecular Diagnostics and Genome Sequencing\nUsing a Repurposed 3D Printer for RNA Extraction","volume":"7","author":"Vandelannoote","year":"2021","journal-title":"ACS Biomater. Sci. Eng."},{"key":"2026081708293629400_cit39","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2020.113972","article-title":"Optimization\nand Clinical Validation of Dual-Target RT-LAMP for SARS-CoV-2","volume":"286","author":"Mohon","year":"2020","journal-title":"J. Virol. Methods"},{"issue":"61","key":"2026081708293629400_cit40","doi-asserted-by":"publisher","first-page":"56205","DOI":"10.1039\/C6RA07529K","article-title":"Rapid Amplification\nof Mycobacterium tuberculosis DNA on\na Paper Substrate","volume":"6","author":"Shetty","year":"2016","journal-title":"RSC Adv."},{"issue":"17","key":"2026081708293629400_cit41","doi-asserted-by":"publisher","first-page":"3082","DOI":"10.1039\/c2lc40423k","article-title":"A Paper\nand Plastic Device for Performing Recombinase\nPolymerase Amplification of HIV DNA","volume":"12","author":"Rohrman","year":"2012","journal-title":"Lab Chip"},{"issue":"4","key":"2026081708293629400_cit42","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0266703","article-title":"A Rapid, Specific,\nExtraction-Less, and Cost-Effective RT-LAMP Test for the Detection\nof SARS-CoV-2 in Clinical Specimens","volume":"17","author":"Marino","year":"2022","journal-title":"PLoS One"},{"issue":"1","key":"2026081708293629400_cit43","doi-asserted-by":"publisher","first-page":"307","DOI":"10.1111\/1751-7915.13737","article-title":"Loop-Mediated\nIsothermal Amplification for the Detection of SARS-CoV-2 in Saliva","volume":"14","author":"Jani\u0301kova\u0301","year":"2021","journal-title":"Microb. Biotechnol."},{"key":"2026081708293629400_cit44","volume-title":"Aerosol Measurement:\nPrinciples, Techniques, and Applications","author":"Baron","year":"2005","edition":"2"}],"container-title":["ACS Omega"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acsomega.4c05784","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/acsodf\/article-pdf\/9\/39\/40832\/3334165\/ao4c05784.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/acsodf\/article-pdf\/9\/39\/40832\/3334165\/ao4c05784.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T12:29:47Z","timestamp":1786969787000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/acsodf\/article\/9\/39\/40832\/357947\/Optimization-of-Reverse-Transcription-Loop"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,9,20]]},"references-count":44,"journal-issue":{"issue":"39","published-online":{"date-parts":[[2024,9,20]]},"published-print":{"date-parts":[[2024,10,1]]}},"URL":"https:\/\/doi.org\/10.1021\/acsomega.4c05784","relation":{},"ISSN":["2470-1343"],"issn-type":[{"value":"2470-1343","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,9,20]]}}}