{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,30]],"date-time":"2026-06-30T00:22:14Z","timestamp":1782778934640,"version":"3.54.5"},"reference-count":18,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,10,13]],"date-time":"2021-10-13T00:00:00Z","timestamp":1634083200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,10,13]],"date-time":"2021-10-13T00:00:00Z","timestamp":1634083200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>This study aimed to develop a highly sensitive SARS-CoV-2 nucleocapsid antigen assay using the single molecule array (Simoa) technology and compare it with real time RT-PCR as used in routine clinical practice with the ambition to achieve a comparative technical and clinical sensitivity. Samples were available from 148 SARS-CoV-2 real time RT-PCR positive and 73 SARS-CoV-2 real time RT-PCR negative oropharyngeal swabs. For determination of technical sensitivity SARS-CoV-2 virus culture material was used. The samples were treated with lysis buffer and analyzed using both an in-house and a pre-commercial SARS-CoV-2 nucleocapsid antigen assay on Simoa. Both nucleocapsid antigen assays have a technical sensitivity corresponding to around 100 SARS-CoV-2 RNA molecules\/mL. Using a cut-off at 0.1\u00a0pg\/mL the pre-commercial SARS-CoV-2 nucleocapsid antigen assay had a sensitivity of 96% (95% CI 91.4\u201398.5%) and specificity of 100% (95% CI 95.1\u2013100%). In comparison the in-house nucleocapsid antigen assay had sensitivity of 95% (95% CI 89.3\u201398.1%) and a specificity of 100% (95% CI 95.1\u2013100%) using a cut-off at 0.01\u00a0pg\/mL. The two SARS-CoV-2 nucleocapsid antigen assays correlated with r\u2009=\u20090.91 (P\u2009&lt;\u20090.0001). The in-house and the pre-commercial SARS-CoV-2 nucleocapsid antigen assay demonstrated technical and clinical sensitivity comparable to real-time RT-PCR methods for identifying SARS-CoV-2 infected patients and thus can be used clinically as well as serve as a reference method for antigen Point of Care Testing.<\/jats:p>","DOI":"10.1038\/s41598-021-99807-7","type":"journal-article","created":{"date-parts":[[2021,10,13]],"date-time":"2021-10-13T17:43:34Z","timestamp":1634147014000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Quantifying SARS-CoV-2 nucleocapsid antigen in oropharyngeal swabs using single molecule array technology"],"prefix":"10.1038","volume":"11","author":[{"given":"Dorte Aa.","family":"Olsen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claus L.","family":"Brasen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S\u00f8ren","family":"Kahns","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeppe B.","family":"Madsen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helene","family":"Kierkegaard","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Henry","family":"Christensen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anders","family":"Jensen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas V.","family":"Sydenham","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jens K.","family":"M\u00f8ller","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonna S.","family":"Madsen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ivan","family":"Brandslund","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,10,13]]},"reference":[{"issue":"8","key":"99807_CR1","doi-asserted-by":"publisher","first-page":"727","DOI":"10.1056\/NEJMoa2001017","volume":"382","author":"N Zhu","year":"2020","unstructured":"Zhu, N. et al. A Novel coronavirus from patients with pneumonia in China. N. Engl. J. Med. 382(8), 727\u2013733 (2020).","journal-title":"N. Engl. J. Med."},{"key":"99807_CR2","unstructured":"World Health Organization. Laboratory testing for coronavirus disease (COVID-19) in suspected human cases. Interim guidance. 19 March 2020."},{"issue":"6","key":"99807_CR3","doi-asserted-by":"publisher","first-page":"595","DOI":"10.1038\/nbt.1641","volume":"28","author":"DM Rissin","year":"2010","unstructured":"Rissin, D. M. et al. Single-molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrations. Nat. Biotechnol. 28(6), 595\u2013599 (2010).","journal-title":"Nat. Biotechnol."},{"issue":"6","key":"99807_CR4","doi-asserted-by":"publisher","first-page":"2279","DOI":"10.1021\/ac103161b","volume":"83","author":"DM Rissin","year":"2011","unstructured":"Rissin, D. M. et al. Simultaneous detection of single molecules and singulated ensembles of molecules enables immunoassays with broad dynamic range. Anal. Chem. 83(6), 2279\u20132285 (2011).","journal-title":"Anal. Chem."},{"issue":"2","key":"99807_CR5","doi-asserted-by":"publisher","first-page":"390","DOI":"10.3390\/s21020390","volume":"21","author":"B Mojsoska","year":"2021","unstructured":"Mojsoska, B. et al. Rapid SARS-CoV-2 detection using electrochemical immunosensor. Sensors (Basel) 21(2), 390 (2021).","journal-title":"Sensors (Basel)"},{"key":"99807_CR6","doi-asserted-by":"publisher","first-page":"1562","DOI":"10.1093\/clinchem\/hvaa213","volume":"66","author":"AF Ogata","year":"2020","unstructured":"Ogata, A. F. et al. Ultra-sensitive serial profiling of SARS-CoV-2 antigens and antibodies in plasma to understand disease progression in COVID-19 patients with severe disease. Clin. Chem. 66, 1562\u20131572 (2020).","journal-title":"Clin. Chem."},{"key":"99807_CR7","volume":"59","author":"NR Pollock","year":"2021","unstructured":"Pollock, N. R. et al. Correlation of SARS-CoV-2 nucleocapsid antigen and RNA concentrations in nasopharyngeal samples from children and adults using an ultrasensitive and quantitative antigen assay. J. Clin. Microbiol. 59, e03077 (2021).","journal-title":"J. Clin. Microbiol."},{"key":"99807_CR8","doi-asserted-by":"publisher","DOI":"10.1101\/2020081420175356","author":"D Shan","year":"2020","unstructured":"Shan, D. et al. SARS-Coronavirus-2 nucleocapsid protein measured in blood using a Simoa ultra-sensitive immunoassay differentiates COVID-19 infection with high clinical sensitivity. Chemrxiv. https:\/\/doi.org\/10.1101\/2020081420175356 (2020).","journal-title":"Chemrxiv."},{"key":"99807_CR9","doi-asserted-by":"publisher","DOI":"10.26434\/chemrxiv12709538v2","author":"D Cate","year":"2021","unstructured":"Cate, D. et al. Antibody screening results for anti-nucleocapsid antibodies towards the development of a SARS-CoV-2 nucleocapsid protein antigen detecting lateral flow assay. ChemRxiv. https:\/\/doi.org\/10.26434\/chemrxiv12709538v2 (2021).","journal-title":"ChemRxiv."},{"key":"99807_CR10","first-page":"1","volume":"181","author":"W Gerhardt","year":"1986","unstructured":"Gerhardt, W. & Keller, H. Evaluation of test data from clinical studies. I. Terminology, graphic interpretation, diagnostic strategies, and selection of sample groups. II. Critical review of the concepts of efficiency, receiver operated characteristics (ROC), and likelihood ratios. Scand. J. Clin. Lab. Investig. Suppl. 181, 1\u201374 (1986).","journal-title":"Scand. J. Clin. Lab. Investig. Suppl."},{"key":"99807_CR11","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1016\/j.jim.2018.05.002","volume":"459","author":"DA Olsen","year":"2018","unstructured":"Olsen, D. A., Kjaer, I. M. & Brandslund, I. Development of a three-plex single molecule immunoassay enabling measurement of the EGFR ligands amphiregulin, betacellulin and transforming growth factor alpha simultaneously in human serum samples. J. Immunol. Methods. 459, 63\u201369 (2018).","journal-title":"J. Immunol. Methods."},{"issue":"10","key":"99807_CR12","doi-asserted-by":"publisher","first-page":"1655","DOI":"10.1515\/cclm-2015-1195","volume":"54","author":"J Kuhle","year":"2016","unstructured":"Kuhle, J. et al. Comparison of three analytical platforms for quantification of the neurofilament light chain in blood samples: ELISA, electrochemiluminescence immunoassay and Simoa. Clin. Chem. Lab. Med. 54(10), 1655\u20131661 (2016).","journal-title":"Clin. Chem. Lab. Med."},{"key":"99807_CR13","doi-asserted-by":"publisher","DOI":"10.1007\/s11427-020-1830-8","author":"B Su","year":"2020","unstructured":"Su, B. et al. Quantification of SARS-CoV-2 antigen levels in the blood of patients with COVID-19. Sci. China Life Sci. https:\/\/doi.org\/10.1007\/s11427-020-1830-8 (2020).","journal-title":"Sci. China Life Sci."},{"key":"99807_CR14","doi-asserted-by":"publisher","DOI":"10.1101\/2020100120203836","author":"LJ Kr\u00fcger","year":"2020","unstructured":"Kr\u00fcger, L. J. et al. Evaluation of the accuracy, ease of use and limit of detection of novel, rapid, antigen-detecting point-of-care diagnostics for SARS-CoV-2. medRxiv. https:\/\/doi.org\/10.1101\/2020100120203836 (2020).","journal-title":"medRxiv."},{"key":"99807_CR15","doi-asserted-by":"publisher","DOI":"10.1016\/jeclinm2020100677","author":"H Gremmelsa","year":"2020","unstructured":"Gremmelsa, H. et al. Real-life validation of the PanbioTM COVID-19 antigen rapid test (Abbott) in community-dwelling subjects with symptoms of potential SARS-CoV-2 infection. EClinicalMedicine. https:\/\/doi.org\/10.1016\/jeclinm2020100677 (2020).","journal-title":"EClinicalMedicine."},{"key":"99807_CR16","doi-asserted-by":"publisher","DOI":"10.1002\/14651858.CD013705","author":"J Dinnes","year":"2020","unstructured":"Dinnes, J. et al. Rapid, point-of-care antigen and molecular-based tests for diagnosis of SARS-CoV-2 infection. Cochrane Database Syst. Rev. https:\/\/doi.org\/10.1002\/14651858.CD013705 (2020).","journal-title":"Cochrane Database Syst. Rev."},{"key":"99807_CR17","doi-asserted-by":"publisher","DOI":"10.1101\/2021012221250042","author":"KK Jakobsen","year":"2021","unstructured":"Jakobsen, K. K. et al. Detection of SARS-CoV-2 infection by rapid antigen test in comparison with RT-PCR in a public setting. MedRxiv. https:\/\/doi.org\/10.1101\/2021012221250042 (2021).","journal-title":"MedRxiv."},{"key":"99807_CR18","doi-asserted-by":"publisher","DOI":"10.1515\/cclm-2021-0694","author":"CL Brasen","year":"2021","unstructured":"Brasen, C. L. et al. Daily monitoring of viral load measured as SARS-CoV-2 antigen and RNA in blood, IL-6, CRP and complement C3d predicts outcome in patients hospitalized with COVID-19. Clin. Chem. Lab. Med. https:\/\/doi.org\/10.1515\/cclm-2021-0694 (2021).","journal-title":"Clin. Chem. Lab. Med."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-99807-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-99807-7","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-99807-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,3]],"date-time":"2022-12-03T16:42:31Z","timestamp":1670085751000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-99807-7"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,10,13]]},"references-count":18,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["99807"],"URL":"https:\/\/doi.org\/10.1038\/s41598-021-99807-7","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,10,13]]},"assertion":[{"value":"29 April 2021","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"1 October 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"13 October 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"20323"}}