An Ultrafast One-Step qRT-PCR Assay for Detection of SARS-CoV-2

An Ultrafast One-Step qRT-PCR Assay for Detection of SARS-CoV-2 | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Method Article An Ultrafast One-Step qRT-PCR Assay for Detection of SARS-CoV-2 Jadranka Milosevic, Mengrou Lu, Wallace Greene, Hong-Zhang He, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-582897/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract We developed an ultrafast one-step RT-qPCR assay for SARS-CoV-2 detection, which can be completed in only 30 mins on benchtop Bio-Rad CFX96. The assay significantly reduces the running time of conventional RT-qPCR: reduced RT step from 10 to 1 min, and reduced PCR cycle of denaturation from 10 to 1 s and extension from 30 to 1 s. A cohort of 60 nasopharyngeal swab samples testing showed that the assay had a clinical sensitivity of 100% and a clinical specificity of 100%. Analytical Biochemistry Ultrafast one-step RT-qPCR assay SARS-CoV-2 detection Full Text Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-582897","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Method Article","associatedPublications":[],"authors":[{"id":30647012,"identity":"6d9c3acc-2f71-4db0-b0ed-a107f7db25a2","order_by":0,"name":"Jadranka Milosevic","email":"","orcid":"","institution":"Captis Diagnostics","correspondingAuthor":false,"prefix":"","firstName":"Jadranka","middleName":"","lastName":"Milosevic","suffix":""},{"id":30647013,"identity":"bad644df-e94c-44cf-b989-16980cdff3ec","order_by":1,"name":"Mengrou Lu","email":"","orcid":"","institution":"Carnegie Mellon University","correspondingAuthor":false,"prefix":"","firstName":"Mengrou","middleName":"","lastName":"Lu","suffix":""},{"id":30647014,"identity":"4d0bf3c9-249f-4067-b330-218cca54cf85","order_by":2,"name":"Wallace Greene","email":"","orcid":"","institution":"Penn State Hershey Medical Center and Penn State College of Medicine","correspondingAuthor":false,"prefix":"","firstName":"Wallace","middleName":"","lastName":"Greene","suffix":""},{"id":30647015,"identity":"2789d55d-6449-4cb5-ab25-a87d8920994a","order_by":3,"name":"Hong-Zhang He","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA1UlEQVRIiWNgGAWjYDACZiBmbGCQA3MSwFwitRiDKeK0MEC0JDaAKWK0yLdzJz4u3GGT3t9+9viDB3+sExvYDx/dgE+LwWHezcYzz6TlzjiTl9iQ2JZuzMCTlnYDrxZm3m3SvG2Hczcw5Bg2JDYclmOQ4DHDq0W+Gazlf7oB/xvDhoQ/h3kIamE4DNZyIMFAAmhLAhsRtoD9wtuWbDjjxrvEGSC/sBHyi3z/2Y2Pedvs5Pn7cw98/AEMsX72w8fwOwwBeCAUG5HKkbSMglEwCkbBKEAHALaGSH8jL7hWAAAAAElFTkSuQmCC","orcid":"","institution":"Captis Diagnostics","correspondingAuthor":true,"prefix":"","firstName":"Hong-Zhang","middleName":"","lastName":"He","suffix":""},{"id":30647066,"identity":"eceebdb4-7524-4d14-958f-d1d2d9302e2d","order_by":4,"name":"Si-Yang Zheng","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA9ElEQVRIiWNgGAWjYDACZgglB8QGCFEeIrQYk6AFChIbiNYi385j+LngV116/+zmbR8+tjHI685IYHzwtg23FoPDPMbSM/vYcmfcOVY8c2Ybg+G2GwnMhnPxaWHmMZDm7eHJbbiRY8zMu40hwexGAps0Lx4t8s08xr95eyTS5UFa/kK0sP/Gp4XhMI+ZNM8PgwQDkBZGqC3M+LQYHGYrs+ZtSDDceCOtmLH3n4ThtjMPmyXnnMPjsP7Dm2/z/KmTl7uRvJnhxxkbebPjyQc/vCnD4zAGDgMGRoQzJICYsQGfeiBgf8DA8IeAmlEwCkbBKBjZAADjg00YYa67CQAAAABJRU5ErkJggg==","orcid":"","institution":"Carnegie Mellon University","correspondingAuthor":true,"prefix":"","firstName":"Si-Yang","middleName":"","lastName":"Zheng","suffix":""}],"badges":[],"createdAt":"2021-06-02 05:12:10","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":true,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false,"coiExplicitlySet":false},"doi":"10.21203/rs.3.rs-582897/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-582897/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":13639611,"identity":"8a83eacc-c75f-48ef-b0cd-84a55a4a3cf2","added_by":"auto","created_at":"2021-09-17 08:56:27","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":412169,"visible":true,"origin":"","legend":"","description":"","filename":"COVID19detectionMSbioRxiv.pdf","url":"https://assets-eu.researchsquare.com/files/rs-582897/v1_covered.pdf"},{"id":9901233,"identity":"8cc090fe-246d-4fd3-afba-9998fbfc58b0","added_by":"auto","created_at":"2021-06-02 17:22:49","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":408749,"visible":true,"origin":"","legend":"","description":"","filename":"COVID19detectionMSbioRxiv.pdf","url":"https://assets-eu.researchsquare.com/files/rs-582897/v1_covered.pdf"}],"financialInterests":"","formattedTitle":"\u003cp\u003eAn Ultrafast One-Step qRT-PCR Assay for Detection of SARS-CoV-2\u003c/p\u003e","fulltext":[{"header":"Full Text","content":"This preprint is available for \u003ca href='/article/rs-582897/latest.pdf' target='_blank'\u003edownload as a PDF\u003c/a\u003e."}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"Carneige Mellon University and Captis Diagnostics Inc","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Ultrafast, one-step RT-qPCR assay, SARS-CoV-2 detection","lastPublishedDoi":"10.21203/rs.3.rs-582897/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-582897/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eWe developed an ultrafast one-step RT-qPCR assay for SARS-CoV-2 detection, which can be completed in only 30 mins on benchtop \u003cem\u003eBio-Rad\u003c/em\u003e CFX96. The assay significantly reduces the running time of conventional RT-qPCR: reduced RT step from 10 to 1 min, and reduced PCR cycle of denaturation from 10 to 1 s and extension from 30 to 1 s. A cohort of 60 nasopharyngeal swab samples testing showed that the assay had a clinical sensitivity of 100% and a clinical specificity of 100%.\u0026nbsp;\u003c/p\u003e\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","manuscriptTitle":"An Ultrafast One-Step qRT-PCR Assay for Detection of SARS-CoV-2","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-06-02 17:22:44","doi":"10.21203/rs.3.rs-582897/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"ae710749-fdbc-4dc9-bbdf-0c2d9df41d3c","owner":[],"postedDate":"June 2nd, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":4734551,"name":"Analytical Biochemistry"}],"tags":[],"updatedAt":"2021-06-02T17:22:44+00:00","versionOfRecord":[],"versionCreatedAt":"2021-06-02 17:22:44","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-582897","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-582897","identity":"rs-582897","version":["v1"]},"buildId":"J0_U0BvcaRcwD8yVFaRlm","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}