{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,16]],"date-time":"2026-05-16T03:44:59Z","timestamp":1778903099189,"version":"3.51.4"},"reference-count":37,"publisher":"Microbiology Society","issue":"9","license":[{"start":{"date-parts":[[2020,9,1]],"date-time":"2020-09-01T00:00:00Z","timestamp":1598918400000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"crossref","award":["APP1174555"],"award-info":[{"award-number":["APP1174555"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"crossref","award":["APP1105905"],"award-info":[{"award-number":["APP1105905"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"crossref","award":["APP1105525"],"award-info":[{"award-number":["APP1105525"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"crossref","award":["GNT2002317"],"award-info":[{"award-number":["GNT2002317"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,9,1]]},"abstract":"<jats:p>\n                    <jats:bold>Introduction.<\/jats:bold>\n                    The SARS-CoV-2 pandemic of 2020 has resulted in unparalleled requirements for RNA extraction kits and enzymes required for virus detection, leading to global shortages. This has necessitated the exploration of alternative diagnostic options to alleviate supply chain issues.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Aim.<\/jats:bold>\n                    To establish and validate a reverse transcription loop-mediated isothermal amplification (RT- LAMP) assay for the detection of SARS-CoV-2 from nasopharyngeal swabs.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Methodology.<\/jats:bold>\n                    We used a commercial RT-LAMP mastermix from OptiGene in combination with a primer set designed to detect the CDC N1 region of the SARS-CoV-2 nucleocapsid (N) gene. A single-tube, single-step fluorescence assay was implemented whereby 1 \u00b5l of universal transport medium (UTM) directly from a nasopharyngeal swab could be used as template, bypassing the requirement for RNA purification. Amplification and detection could be conducted in any thermocycler capable of holding 65 \u00b0C for 30 min and measure fluorescence in the FAM channel at 1 min intervals.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Results.<\/jats:bold>\n                    Assay evaluation by assessment of 157 clinical specimens previously screened by E-gene RT-qPCR revealed assay sensitivity and specificity of 87 and 100%, respectively. Results were fast, with an average time-to-positive (Tp) for 93 clinical samples of 14 min (\n                    <jats:sc>sd<\/jats:sc>\n                    \u00b17 min). Using dilutions of SARS-CoV-2 virus spiked into UTM, we also evaluated assay performance against FDA guidelines for implementation of emergency-use diagnostics and established a limit-of-detection of 54 Tissue Culture Infectious Dose 50 per ml (TCID\n                    <jats:sub>50<\/jats:sub>\n                    ml\n                    <jats:sup>\u22121<\/jats:sup>\n                    ), with satisfactory assay sensitivity and specificity. A comparison of 20 clinical specimens between four laboratories showed excellent interlaboratory concordance; performing equally well on three different, commonly used thermocyclers, pointing to the robustness of the assay.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Conclusion.<\/jats:bold>\n                    With a simplified workflow, The N1 gene Single Tube Optigene LAMP assay (N1-STOP-LAMP) is a powerful, scalable option for specific and rapid detection of SARS-CoV-2 and an additional resource in the diagnostic armamentarium against COVID-19.\n                  <\/jats:p>","DOI":"10.1099\/jmm.0.001238","type":"journal-article","created":{"date-parts":[[2020,8,5]],"date-time":"2020-08-05T10:15:55Z","timestamp":1596622555000},"page":"1169-1178","source":"Crossref","is-referenced-by-count":73,"title":["Validation of a single-step, single-tube reverse transcription loop-mediated isothermal amplification assay for rapid detection of SARS-CoV-2 RNA"],"prefix":"10.1099","volume":"69","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5751-8503","authenticated-orcid":false,"given":"Jean Y. H.","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Infectious Diseases, Monash Health, Clayton, Victoria, Australia"},{"name":"Department of Microbiology and Immunology, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4555-2517","authenticated-orcid":false,"given":"Nickala","family":"Best","sequence":"additional","affiliation":[{"name":"GenWorks Pty Ltd, Thebarton, South Australia, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2493-3465","authenticated-orcid":false,"given":"Julie","family":"McAuley","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jessica L.","family":"Porter","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Torsten","family":"Seemann","sequence":"additional","affiliation":[{"name":"Microbiological Diagnostic Unit Public Health Laboratory, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"},{"name":"Department of Microbiology and Immunology, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7689-6531","authenticated-orcid":false,"given":"Mark B.","family":"Schultz","sequence":"additional","affiliation":[{"name":"Microbiological Diagnostic Unit Public Health Laboratory, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8782-6128","authenticated-orcid":false,"given":"Michelle","family":"Sait","sequence":"additional","affiliation":[{"name":"Microbiological Diagnostic Unit Public Health Laboratory, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicole","family":"Orlando","sequence":"additional","affiliation":[{"name":"Microbiological Diagnostic Unit Public Health Laboratory, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3957-666X","authenticated-orcid":false,"given":"Karolina","family":"Mercoulia","sequence":"additional","affiliation":[{"name":"Microbiological Diagnostic Unit Public Health Laboratory, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Susan A.","family":"Ballard","sequence":"additional","affiliation":[{"name":"Microbiological Diagnostic Unit Public Health Laboratory, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julian","family":"Druce","sequence":"additional","affiliation":[{"name":"Victorian Infectious Diseases Reference Laboratory, Melbourne Health at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thomas","family":"Tran","sequence":"additional","affiliation":[{"name":"Victorian Infectious Diseases Reference Laboratory, Melbourne Health at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0297-1260","authenticated-orcid":false,"given":"Mike G.","family":"Catton","sequence":"additional","affiliation":[{"name":"Victorian Infectious Diseases Reference Laboratory, Melbourne Health at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Melinda J.","family":"Pryor","sequence":"additional","affiliation":[{"name":"360Biolabs, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huanhuan L.","family":"Cui","sequence":"additional","affiliation":[{"name":"360Biolabs, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angela","family":"Luttick","sequence":"additional","affiliation":[{"name":"360Biolabs, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sean","family":"McDonald","sequence":"additional","affiliation":[{"name":"GenWorks Pty Ltd, Thebarton, South Australia, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Arran","family":"Greenhalgh","sequence":"additional","affiliation":[{"name":"GenWorks Pty Ltd, Thebarton, South Australia, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jason C.","family":"Kwong","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases, Austin Health, Heidelberg, Victoria, Australia"},{"name":"Department of Microbiology and Immunology, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7789-8360","authenticated-orcid":false,"given":"Norelle L.","family":"Sherry","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases, Austin Health, Heidelberg, Victoria, Australia"},{"name":"Microbiological Diagnostic Unit Public Health Laboratory, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1069-4224","authenticated-orcid":false,"given":"Maryza","family":"Graham","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Monash Health, Clayton, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0086-5576","authenticated-orcid":false,"given":"Tuyet","family":"Hoang","sequence":"additional","affiliation":[{"name":"Microbiological Diagnostic Unit Public Health Laboratory, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"},{"name":"Department of Microbiology and Immunology, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4703-5927","authenticated-orcid":false,"given":"Marion","family":"Herisse","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1202-6614","authenticated-orcid":false,"given":"Sacha J.","family":"Pidot","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Deborah A.","family":"Williamson","sequence":"additional","affiliation":[{"name":"Melbourne Health, Melbourne, Victoria, Australia"},{"name":"Microbiological Diagnostic Unit Public Health Laboratory, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"},{"name":"Department of Microbiology and Immunology, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0237-1473","authenticated-orcid":false,"given":"Benjamin P.","family":"Howden","sequence":"additional","affiliation":[{"name":"Microbiological Diagnostic Unit Public Health Laboratory, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"},{"name":"Department of Microbiology and Immunology, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"},{"name":"Department of Infectious Diseases, Austin Health, Heidelberg, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6982-8074","authenticated-orcid":false,"given":"Ian R.","family":"Monk","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0150-123X","authenticated-orcid":false,"given":"Timothy P.","family":"Stinear","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"345","reference":[{"key":"R1","article-title":"Coronavirus disease 2019 (COVID-19) Guidelines for health services and general practitioners - Version 17, 5\/04\/2020. State Government of Victoria, Australia."},{"key":"R2","article-title":"Guidance and standard operating procedure: COVID-19 virus testing in NHS laboratories, 16\/03\/2020. NHS, London","year":"2020"},{"key":"R3","article-title":"A 2019-Novel Coronavirus (2019-nCoV) real-time RT-PCR panel primers and probes, 24\/01\/2020. US Department of Health & Human Services, CDC, Atlanta","year":"2020"},{"key":"R4","article-title":"Real-time RT-PCR panel for detection 2019-Novel Coronavirus, 24\/01\/2020. US Department of Health & Human Services, CDC, Atlanta","year":"2020"},{"key":"R5","article-title":"Policy for diagnostic tests for Coronavirus disease \u2013 2019 during the public health emergency \u2013 Immediately in effect guidance for clinical laboratories, commercial manufacturers, and food and Drug administration staff, 16\/03\/2020. US Department of Health & Human Services, Food and Drug Administration"},{"key":"R6","volume-title":"Laboratory Testing for Coronavirus Disease 2019 (COVID-19) in Suspected Human Cases, 2\/03\/2020","year":"2020"},{"key":"R7","volume-title":"Coronavirus Disease (COVID-19) Technical Guidance: Laboratory Testing for 2019-nCoV in Humans. 3. Molecular Assays to Diagnose COVID-19, 03\/2020","year":"2020"},{"key":"R8","volume-title":"Deputy Chief Medical Officer\u2019s Press Conference About COVID-19 on 16 March","author":"Kelly","year":"2020"},{"key":"R9","doi-asserted-by":"publisher","DOI":"10.1038\/d41591-020-00005-1"},{"key":"R10","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-020-0499-y"},{"key":"R11","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/28.12.e63"},{"key":"R12","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.42.5.1956-1961.2004"},{"key":"R13","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.41.6.2616-2622.2003"},{"key":"R14","doi-asserted-by":"publisher","DOI":"10.1007\/s10156-013-0590-0"},{"key":"R15","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2005.057901"},{"key":"R16","doi-asserted-by":"publisher","DOI":"10.1158\/1078-0432.CCR-06-2512"},{"key":"R17","article-title":"Rapid molecular detection of SARS-CoV-2 (COVID-19) virus RNA using colorimetric lamp","author":"Zhang","year":"2020","journal-title":"medRxiv"},{"key":"R18","article-title":"Rapid colorimetric detection of COVID-19 coronavirus using a reverse tran-scriptional loop-mediated isothermal amplification (RT-LAMP) diagnostic plat-form: iLACO","author":"Yu","year":"2020","journal-title":"medRxiv"},{"key":"R19","doi-asserted-by":"crossref","DOI":"10.1016\/j.cmi.2020.04.001","article-title":"Rapid and visual detection of 2019 novel coronavirus (SARS-CoV-2) by a reverse transcription loop-mediated isothermal amplification assay","author":"Yan","year":"2020","journal-title":"Clin Microbiol Infect"},{"key":"R20","doi-asserted-by":"crossref","DOI":"10.1016\/j.jmoldx.2020.03.006","article-title":"Development of reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays targeting SARS-CoV-2","author":"Park","year":"2020","journal-title":"J Mol Diagnostics"},{"key":"R21","doi-asserted-by":"publisher","DOI":"10.3390\/jcm9030623"},{"key":"R22","doi-asserted-by":"publisher","DOI":"10.2139\/ssrn.3564906"},{"key":"R23","article-title":"Development of a novel reverse transcription loop-mediated isothermal amplification method for rapid detection of SARS-CoV-2","author":"Lu","year":"2020","journal-title":"Virol Sin"},{"key":"R24","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-020-0513-4"},{"key":"R25","doi-asserted-by":"publisher","DOI":"10.5694\/mja2.50569"},{"key":"R26","doi-asserted-by":"publisher","DOI":"10.1038\/srep45199"},{"key":"R27","doi-asserted-by":"publisher","DOI":"10.2807\/1560-7917.ES.2020.25.3.2000045"},{"key":"R28","doi-asserted-by":"publisher","DOI":"10.1080\/22221751.2020.1756698"},{"key":"R29","doi-asserted-by":"publisher","DOI":"10.3390\/mi11030306"},{"key":"R30","article-title":"SARS-CoV-2 diagnostic pipeline, 2020","year":"2020"},{"key":"R31","doi-asserted-by":"publisher","DOI":"10.3201\/eid1306.061572"},{"key":"R32","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0006448"},{"key":"R33","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-40960-5"},{"key":"R34","article-title":"High-surety isothermal amplification and detection of SARS-CoV-2, including with crude enzymes","author":"Bhadra","year":"2020","journal-title":"BioRxiv"},{"key":"R35","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2008.08.001"},{"key":"R36","doi-asserted-by":"publisher","DOI":"10.4142\/jvs.2016.17.3.421"},{"key":"R37","volume-title":"National Statement on Ethical Conduct in Human Research 2007 (Updated 2018). The National Health and Medical Research Council, the Australian Research Council and Universities Australia","year":"2018"}],"container-title":["Journal of Medical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jmm\/10.1099\/jmm.0.001238?crawler=true&mimetype=application\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,12,14]],"date-time":"2023-12-14T18:25:23Z","timestamp":1702578323000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jmm\/10.1099\/jmm.0.001238"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,9,1]]},"references-count":37,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2020,9,1]]}},"URL":"https:\/\/doi.org\/10.1099\/jmm.0.001238","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2020.04.28.067363","asserted-by":"object"}]},"ISSN":["0022-2615","1473-5644"],"issn-type":[{"value":"0022-2615","type":"print"},{"value":"1473-5644","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,9,1]]}}}