Surface‑Engineered Primer Immobilization Enables Simplified and Affordable Nucleic‑Acid Capture for Molecular Diagnostics in Sub‑Saharan Africa

Surface‑Engineered Primer Immobilization Enables Simplified and Affordable Nucleic‑Acid Capture for Molecular Diagnostics in Sub‑Saharan Africa | 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 Research Article Surface‑Engineered Primer Immobilization Enables Simplified and Affordable Nucleic‑Acid Capture for Molecular Diagnostics in Sub‑Saharan Africa Tochukwu Dubem Anyaduba This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9369758/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 Molecular diagnostic (MDx) technologies remain prohibitively expensive for many laboratories in sub-Saharan Africa. this is because current platforms rely on silica-based extraction processes that require multiple wash buffers, magnetic components, and ethanol handling. This study presents a simplified, silica-free approach that uses functionalized polymer surfaces for nucleic-acid capture. Polycarbonate was selected as a model sub-strate based on its thermal stability and innate carbonate groups, which were chemically and optically mod-ified by low-cost acetone and ultraviolet (UV) pretreatments to generate surface carboxyl (–COOH) groups. Quantification using toluidine blue-O dye revealed a twofold increase in COOH density after combined ace-tone–UV treatment, enabling efficient covalent attachment of primers through branched polyethyleneimine linkers. Comparative fluorescence assays confirmed that BPEI-linked surfaces carried approximately 2.6 times more oligonucleotides than ethylenediamine-linked surfaces. Hybridization experiments using fluores-cence resonance energy transfer (FRET) verified specific DNA capture under both phosphate and chaotropic lysis conditions, demonstrating compatibility with standard sample buffers. Although amplification directly from immobilized primers was not performed, hybridization and payload analyses showed that the sur-face contained sufficient primer density for enzymatic reactions. The described process replaces complex silica-based extraction with a single solid-phase capture step, offering a simple, inexpensive, and locally manufacturable pathway toward affordable molecular diagnostics in resource-limited settings. Biotechnology and Bioengineering molecular diagnostics primer payload solid-phase DNA Purification affordable diagnostics global health technology infectious disease sub-Saharan Africa LMIC Low-resource environment frugal diagnostics Full Text Additional Declarations The authors declare no competing interests. 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-9369758","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":620413679,"identity":"b38794f3-4246-4986-a600-6c96201f44a9","order_by":0,"name":"Tochukwu Dubem Anyaduba","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0002-9852-8879","institution":"Hezekiah University Umudi, Imo State, Nigeria","correspondingAuthor":true,"prefix":"","firstName":"Tochukwu","middleName":"Dubem","lastName":"Anyaduba","suffix":""}],"badges":[],"createdAt":"2026-04-09 14:18:48","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-9369758/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9369758/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":106725279,"identity":"6fc0a908-63b3-41fb-9a37-5c4380775aa2","added_by":"auto","created_at":"2026-04-12 18:32:13","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1277432,"visible":true,"origin":"","legend":"","description":"","filename":"DNACaptureonPlasticsTDAnyaduba.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9369758/v1_covered_d0fda772-179e-4d9c-817d-735a85487cae.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003eSurface‑Engineered Primer Immobilization Enables Simplified and Affordable Nucleic‑Acid Capture for Molecular Diagnostics in Sub‑Saharan Africa\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":true,"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":"molecular diagnostics, primer payload, solid-phase, DNA Purification, affordable diagnostics, global health technology, infectious disease, sub-Saharan Africa, LMIC, Low-resource environment, frugal diagnostics","lastPublishedDoi":"10.21203/rs.3.rs-9369758/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9369758/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eMolecular diagnostic (MDx) technologies remain prohibitively expensive for many laboratories in sub-Saharan Africa. this is because current platforms rely on silica-based extraction processes that require multiple wash buffers, magnetic components, and ethanol handling. This study presents a simplified, silica-free approach that uses functionalized polymer surfaces for nucleic-acid capture. Polycarbonate was selected as a model sub-strate based on its thermal stability and innate carbonate groups, which were chemically and optically mod-ified by low-cost acetone and ultraviolet (UV) pretreatments to generate surface carboxyl (\u0026ndash;COOH) groups. Quantification using toluidine blue-O dye revealed a twofold increase in COOH density after combined ace-tone\u0026ndash;UV treatment, enabling efficient covalent attachment of primers through branched polyethyleneimine linkers. Comparative fluorescence assays confirmed that BPEI-linked surfaces carried approximately 2.6 times more oligonucleotides than ethylenediamine-linked surfaces. Hybridization experiments using fluores-cence resonance energy transfer (FRET) verified specific DNA capture under both phosphate and chaotropic lysis conditions, demonstrating compatibility with standard sample buffers. Although amplification directly from immobilized primers was not performed, hybridization and payload analyses showed that the sur-face contained sufficient primer density for enzymatic reactions. 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