{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T02:58:48Z","timestamp":1792637928464,"version":"4.5.0"},"reference-count":32,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,7,14]],"date-time":"2021-07-14T00:00:00Z","timestamp":1626220800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,7,14]],"date-time":"2021-07-14T00:00:00Z","timestamp":1626220800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100001395","name":"Wisconsin Alumni Research Foundation","doi-asserted-by":"publisher","award":["COVID-19 Accelerator Challenge Award"],"award-info":[{"award-number":["COVID-19 Accelerator Challenge Award"]}],"id":[{"id":"10.13039\/100001395","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"U.S. Department of Health & Human Services | National Institutes of Health","doi-asserted-by":"publisher","award":["NIH P30CA014520"],"award-info":[{"award-number":["NIH P30CA014520"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"U.S. Department of Health & Human Services | National Institutes of Health","doi-asserted-by":"publisher","award":["NIH 1R01CA247479"],"award-info":[{"award-number":["NIH 1R01CA247479"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"U.S. Department of Health & Human Services | National Institutes of Health","doi-asserted-by":"publisher","award":["NIH 1R01CA247479-01"],"award-info":[{"award-number":["NIH 1R01CA247479-01"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"U.S. Department of Health & Human Services | National Institutes of Health","doi-asserted-by":"publisher","award":["NIH 5UL1TR002373-03"],"award-info":[{"award-number":["NIH 5UL1TR002373-03"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"U.S. Department of Health & Human Services | National Institutes of Health","doi-asserted-by":"publisher","award":["NIH OD011106"],"award-info":[{"award-number":["NIH OD011106"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"name":"U.S. Department of Health & Human Services | National Institutes of Health"},{"name":"U.S. Department of Health & Human Services | National Institutes of Health"},{"name":"U.S. Department of Health & Human Services | National Institutes of Health"},{"name":"U.S. Department of Health & Human Services | National Institutes of Health"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Nat Commun"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The COVID-19 pandemic exposed difficulties in scaling current quantitative PCR (qPCR)-based diagnostic methodologies for large-scale infectious disease testing. Bottlenecks include lengthy multi-step processes for nucleic acid extraction followed by qPCR readouts, which require costly instrumentation and infrastructure, as well as reagent and plastic consumable shortages stemming from supply chain constraints. Here we report an Oil Immersed Lossless Total Analysis System (OIL-TAS), which integrates RNA extraction and detection onto a single device that is simple, rapid, cost effective, and requires minimal supplies and infrastructure to perform. We validated the performance of OIL-TAS using contrived SARS-CoV-2 viral particle samples and clinical nasopharyngeal swab samples. OIL-TAS showed a 93% positive predictive agreement (n\u2009=\u200957) and 100% negative predictive agreement (n\u2009=\u200910) with clinical SARS-CoV-2 qPCR assays in testing clinical samples, highlighting its potential to be a faster, cheaper, and easier-to-deploy alternative for infectious disease testing.<\/jats:p>","DOI":"10.1038\/s41467-021-24463-4","type":"journal-article","created":{"date-parts":[[2021,7,14]],"date-time":"2021-07-14T06:03:06Z","timestamp":1626242586000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":50,"title":["Oil immersed lossless total analysis system for integrated RNA extraction and detection of SARS-CoV-2"],"prefix":"10.1038","volume":"12","author":[{"given":"Duane S.","family":"Juang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Terry D.","family":"Juang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dawn M.","family":"Dudley","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christina M.","family":"Newman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Molly A.","family":"Accola","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William M.","family":"Rehrauer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9831-6895","authenticated-orcid":false,"given":"Thomas C.","family":"Friedrich","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2139-470X","authenticated-orcid":false,"given":"David H.","family":"O\u2019Connor","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0415-9006","authenticated-orcid":false,"given":"David J.","family":"Beebe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,7,14]]},"reference":[{"key":"24463_CR1","doi-asserted-by":"publisher","first-page":"10","DOI":"10.1038\/s41421-020-0148-0","volume":"6","author":"H Wang","year":"2020","unstructured":"Wang, H. et al. Phase-adjusted estimation of the number of Coronavirus Disease 2019 cases in Wuhan, China. Cell Discov. 6, 10 (2020).","journal-title":"Cell Discov."},{"key":"24463_CR2","doi-asserted-by":"crossref","unstructured":"Hui, D. S. et al. The continuing 2019-nCoV epidemic threat of novel coronaviruses to global health\u2014the latest 2019 novel coronavirus outbreak in Wuhan, China. Int. J. Infect. Dis. 91, 264\u2013266 (2020).","DOI":"10.1016\/j.ijid.2020.01.009"},{"key":"24463_CR3","unstructured":"Moore, K. & Conti, S. Grenova\u2019s plan to stop supply shortages in COVID-19 testing labs. Richmond Grid. https:\/\/richmondgrid.com\/grenovas-plan-to-stop-supply-shortages-in-covid-19-testing-labs\/ (2020)."},{"key":"24463_CR4","unstructured":"Wu, K. J. Coronavirus testing labs again lack key supplies. N. Y. Times https:\/\/www.nytimes.com\/2020\/07\/23\/health\/coronavirus-testing-supplyshortage.html (2020)."},{"key":"24463_CR5","doi-asserted-by":"publisher","first-page":"17065","DOI":"10.1021\/acsami.8b03627","volume":"10","author":"C Li","year":"2018","unstructured":"Li, C. et al. Exclusive liquid repellency: an open multi-liquid-phase technology for rare cell culture and single-cell processing. ACS Appl. Mater. Interfaces 10, 17065\u201317070 (2018).","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"24463_CR6","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1177\/2472630319853219","volume":"24","author":"C Li","year":"2019","unstructured":"Li, C., Niles, D. J., Juang, D. S., Lang, J. M. & Beebe, D. J. Automated system for small-population single-particle processing enabled by exclusive liquid repellency. SLAS Technol. 24, 535\u2013542 (2019).","journal-title":"SLAS Technol."},{"key":"24463_CR7","doi-asserted-by":"publisher","first-page":"1747","DOI":"10.1039\/c1lc00004g","volume":"11","author":"SM Berry","year":"2011","unstructured":"Berry, S. M., Alarid, E. T. & Beebe, D. J. One-step purification of nucleic acid for gene expression analysis via immiscible filtration assisted by surface tension (IFAST). Lab Chip 11, 1747\u20131753 (2011).","journal-title":"Lab Chip"},{"key":"24463_CR8","doi-asserted-by":"publisher","first-page":"1033","DOI":"10.1007\/s10544-011-9573-z","volume":"13","author":"SM Berry","year":"2011","unstructured":"Berry, S. M., Strotman, L. N., Kueck, J. D., Alarid, E. T. & Beebe, D. J. Purification of cell subpopulations via immiscible filtration assisted by surface tension (IFAST). Biomed. Microdevices 13, 1033\u20131042 (2011).","journal-title":"Biomed. Microdevices"},{"key":"24463_CR9","doi-asserted-by":"publisher","first-page":"206","DOI":"10.1177\/2211068212462023","volume":"18","author":"SM Berry","year":"2013","unstructured":"Berry, S. M., Regehr, K. J., Casavant, B. P. & Beebe, D. J. Automated operation of immiscible filtration assisted by surface tension (IFAST) arrays for streamlined analyte isolation. J. Lab. Autom. 18, 206\u2013211 (2013).","journal-title":"J. Lab. Autom."},{"key":"24463_CR10","doi-asserted-by":"publisher","first-page":"391","DOI":"10.1039\/C2LC41136A","volume":"13","author":"BP Casavant","year":"2013","unstructured":"Casavant, B. P. et al. The VerIFAST: an integrated method for cell isolation and extracellular\/intracellular staining. Lab Chip 13, 391\u2013396 (2013).","journal-title":"Lab Chip"},{"key":"24463_CR11","doi-asserted-by":"publisher","first-page":"e0143631","DOI":"10.1371\/journal.pone.0143631","volume":"10","author":"SM Berry","year":"2015","unstructured":"Berry, S. M. et al. Using exclusion-based sample preparation (ESP) to reduce viral load assay cost. PLoS ONE 10, e0143631 (2015).","journal-title":"PLoS ONE"},{"key":"24463_CR12","doi-asserted-by":"publisher","first-page":"11848","DOI":"10.1021\/acs.analchem.9b02572","volume":"91","author":"DS Juang","year":"2019","unstructured":"Juang, D. S., Berry, S. M., Li, C., Lang, J. M. & Beebe, D. J. Centrifugation-assisted immiscible fluid filtration for dual-bioanalyte extraction. Anal. Chem. 91, 11848\u201311855 (2019).","journal-title":"Anal. Chem."},{"key":"24463_CR13","doi-asserted-by":"publisher","first-page":"E63","DOI":"10.1093\/nar\/28.12.e63","volume":"28","author":"T Notomi","year":"2000","unstructured":"Notomi, T. et al. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 28, E63 (2000).","journal-title":"Nucleic Acids Res."},{"key":"24463_CR14","doi-asserted-by":"publisher","first-page":"59","DOI":"10.2144\/000114253","volume":"58","author":"NA Tanner","year":"2015","unstructured":"Tanner, N. A., Zhang, Y. & Evans, T. C. Jr Visual detection of isothermal nucleic acid amplification using pH-sensitive dyes. Biotechniques 58, 59\u201368 (2015).","journal-title":"Biotechniques"},{"key":"24463_CR15","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1016\/j.ab.2013.10.038","volume":"447","author":"SM Berry","year":"2014","unstructured":"Berry, S. M. et al. Weak protein\u2013protein interactions revealed by immiscible filtration assisted by surface tension. Anal. Biochem. 447, 133\u2013140 (2014).","journal-title":"Anal. Biochem."},{"key":"24463_CR16","doi-asserted-by":"publisher","first-page":"6355","DOI":"10.1021\/ac500574t","volume":"86","author":"BP Casavant","year":"2014","unstructured":"Casavant, B. P., Guckenberger, D. J., Beebe, D. J. & Berry, S. M. Efficient sample preparation from complex biological samples using a sliding lid for immobilized droplet extractions. Anal. Chem. 86, 6355\u20136362 (2014).","journal-title":"Anal. Chem."},{"key":"24463_CR17","doi-asserted-by":"publisher","first-page":"15040","DOI":"10.1021\/acsami.6b02555","volume":"8","author":"SM Berry","year":"2016","unstructured":"Berry, S. M. et al. AirJump: using interfaces to instantly perform simultaneous extractions. ACS Appl. Mater. Interfaces 8, 15040\u201315045 (2016).","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"24463_CR18","unstructured":"MagNA Pure 96 Instrument. High-throughput Robotic Workstation for Fully Automated Purification of Nucleic Acids from up to 96 Samples. Roche Molecular Systems Inc. https:\/\/lifescience.roche.com\/en_us\/products\/magna-pure-96-instrument-382411-1.html#overview (2020)."},{"key":"24463_CR19","unstructured":"Applied Biosystems\u2122 MagMAXTM-96 Viral RNA Isolation Kit. Thermo Fisher Scientific Inc., https:\/\/www.thermofisher.com\/order\/catalog\/product\/AM1836#\/AM1836 (2017)."},{"key":"24463_CR20","unstructured":"Magnetic Bead-based Isolation of Viral RNA\/DNA and Bacterial DNA\u2014NucleoMag Pathogen. Takara Bio Inc., https:\/\/www.takarabio.com\/products\/nucleic-acid-purification\/viral-dna-and-rna-purification-kits\/nucleomag-pathogen (2017)."},{"key":"24463_CR21","doi-asserted-by":"publisher","first-page":"495","DOI":"10.1128\/jcm.28.3.495-503.1990","volume":"28","author":"R Boom","year":"1990","unstructured":"Boom, R. et al. Rapid and simple method for purification of nucleic acids. J. Clin. Microbiol. 28, 495\u2013503 (1990).","journal-title":"J. Clin. Microbiol."},{"key":"24463_CR22","doi-asserted-by":"crossref","unstructured":"Escobar, M. D. & Hunt, J. L. A cost-effective RNA extraction technique from animal cells and tissue using silica columns. J. Biol. Methods 4, e72 (2017).","DOI":"10.14440\/jbm.2017.184"},{"key":"24463_CR23","doi-asserted-by":"publisher","DOI":"10.1038\/srep45199","volume":"7","author":"H He","year":"2017","unstructured":"He, H. et al. Integrated DNA and RNA extraction using magnetic beads from viral pathogens causing acute respiratory infections. Sci. Rep. 7, 45199 (2017).","journal-title":"Sci. Rep."},{"key":"24463_CR24","doi-asserted-by":"publisher","first-page":"e3000107","DOI":"10.1371\/journal.pbio.3000107","volume":"17","author":"P Oberacker","year":"2019","unstructured":"Oberacker, P. et al. Bio-On-Magnetic-Beads (BOMB): open platform for high-throughput nucleic acid extraction and manipulation. PLoS Biol. 17, e3000107 (2019).","journal-title":"PLoS Biol."},{"key":"24463_CR25","doi-asserted-by":"publisher","DOI":"10.1186\/1756-0500-5-45","volume":"5","author":"H Yaffe","year":"2012","unstructured":"Yaffe, H. et al. LogSpin: a simple, economical and fast method for RNA isolation from infected or healthy plants and other eukaryotic tissues. BMC Res. Notes 5, 45 (2012).","journal-title":"BMC Res. Notes"},{"key":"24463_CR26","doi-asserted-by":"publisher","unstructured":"Scallan, M. F. et al. Validation of a Lysis Buffer containing 4\u2009M guanidinium thiocyanate (GITC)\/triton X-100 for extraction of SARS-CoV-2 RNA for COVID-19 testing: comparison of formulated lysis buffers containing 4 to 6\u2009M GITC, Roche external lysis buffer and qiagen RTL lysis buffer. Preprint at bioRxiv https:\/\/doi.org\/10.1101\/2020.04.05.026435 (2020).","DOI":"10.1101\/2020.04.05.026435"},{"key":"24463_CR27","doi-asserted-by":"crossref","unstructured":"Broughton, J. P. et al. CRISPR\u2013Cas12-based detection of SARS-CoV-2. Nat. Biotechnol. 38, 870\u2013874 (2020).","DOI":"10.1038\/s41587-020-0513-4"},{"key":"24463_CR28","unstructured":"Broughton, J. P. et al. A Protocol For Rapid Detection of the 2019 Novel Coronavirus SARS-CoV-2 Using CRISPR Diagnostics: SARS-CoV-2 DETECTR. Mammoth Biosciences Inc., (2020)."},{"key":"24463_CR29","unstructured":"Color Genomics SARS-CoV-2 LAMP Diagnostic Assay Version 1.2. Color Genomics Inc., https:\/\/www.color.com\/wp-content\/uploads\/2020\/05\/Color-LAMP-Diagnostic-Assay_v1-2_Updated-052120.pdf (2020)."},{"key":"24463_CR30","doi-asserted-by":"crossref","unstructured":"Rabe, B. A. & Cepko, C. SARS-CoV-2 detection using isothermal amplification and a rapid, inexpensive protocol for sample inactivation and purification. Proc. Natl. Acad. Sci. U. S. A. 117, 24450\u201324458 (2020).","DOI":"10.1073\/pnas.2011221117"},{"key":"24463_CR31","doi-asserted-by":"publisher","first-page":"91","DOI":"10.1016\/j.jviromet.2018.02.012","volume":"255","author":"KA Curtis","year":"2018","unstructured":"Curtis, K. A. et al. A multiplexed RT-LAMP assay for detection of group M HIV-1 in plasma or whole blood. J. Virol. Methods 255, 91\u201397 (2018).","journal-title":"J. Virol. Methods"},{"key":"24463_CR32","unstructured":"BEI Certificate of Analysis for product NR-52286 SARS-Related Coronavirus 2, Isolate USAWA1\/2020, Heat Inactivated. BEI Resources, https:\/\/www.beiresources.org\/Catalog\/antigen\/NR-52286.aspx (2020)."}],"container-title":["Nature Communications"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41467-021-24463-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41467-021-24463-4","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41467-021-24463-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,2]],"date-time":"2022-12-02T08:30:14Z","timestamp":1669969814000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41467-021-24463-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,14]]},"references-count":32,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["24463"],"URL":"https:\/\/doi.org\/10.1038\/s41467-021-24463-4","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2020.09.30.20204842","asserted-by":"object"}]},"ISSN":["2041-1723"],"issn-type":[{"value":"2041-1723","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,7,14]]},"assertion":[{"value":"30 September 2020","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 June 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"14 July 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare the following competing interests. David J. Beebe holds equity in Bellbrook Labs LLC, Tasso Inc., Salus Discovery LLC, Lynx Biosciences Inc., Stacks to the Future LLC, Turba LLC, Onexio Biosystems LLC, and Flambeau Diagnostics LLC. David J. Beebe is also a consultant for Abbott Laboratories. Duane S. Juang holds equity in Turba LLC. A patent application (16\/894273) of which D.J.B., D.S.J., T.D.J., D.H.O., and T.C.F. are inventors was filed through the Wisconsin Alumni Research Foundation for the OIL-TAS technology described in this work. All other authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"4317"}}