Cocktail-Inspired All-in-One Magnetofluidic Cartridge Platform for Versatile Nucleic Acid Testing

Cocktail-Inspired All-in-One Magnetofluidic Cartridge Platform for Versatile Nucleic Acid Testing | 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 Article Cocktail-Inspired All-in-One Magnetofluidic Cartridge Platform for Versatile Nucleic Acid Testing Nan Li, Tianping Zhou, Changding Li, Xiaoye Huo, Guangyang Gou, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8883177/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 Low-cost, portable, and high-performance nucleic acid detection technologies are essential for point-of-care diagnosis of infectious diseases. Here, we report an ultralow-cost (~ $ 0.1) magnetofluidic cartridge system featuring a novel tapered cylindrical-like cartridge with stable vertical separation of multiple reagents using a combination of silicone oil and wax layers in a cocktail-like configuration. We systematically investigate the mechanisms that enable shake-proof vertical isolation of multiple immiscible reagent layers, independent of aqueous reagent density, thereby allowing the stable maintenance of arbitrary numbers and arrangements of reagent layers. This innovation facilitates fully automated magnetic bead-based nucleic acid extraction coupled with multiplex PCR or CRISPR‒Cas-assisted isothermal detection in a handheld analyser. The unique reagent isolation in the cartridge overcomes the limitations of previous magnetofluidic systems by physically separating elution and amplification reagents, allowing broader assay compatibility. SARS-CoV-2 and influenza A/B detection reached 0.1 copies/µL sensitivity after 60 minutes (PCR) and 30 minutes (CRISPR). Clinical testing of 33 samples showed 100% concordance with standard methods. This low-cost integrated cartridge system offers a robust platform for rapid, sensitive, and multiplexed nucleic acid testing in resource-limited settings. Physical sciences/Nanoscience and technology/Nanobiotechnology/Microfluidics Physical sciences/Nanoscience and technology/Nanobiotechnology/Biosensors magnetofluidic nucleic acid detection point-of-care testing PCR CRISPR Full Text Additional Declarations There is no conflict of interest Supplementary Files VideoS4.mp4 Video S4 Shake-proof test of paraffin wax layer stability in cartridge SupplementaryMaterial.docx Supplementary information VideoS3.mp4 Video S3 Vibration platform test of oil-water phase separation stability VideoS2.mp4 Video S2 Automated processing on type B cartridge for one-pot RT-RPA-CRISPR VideoS1.mp4 Video S1 Automated processing on type A cartridge for multiplex PCR 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. 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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-8883177","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":599924815,"identity":"ad51c4fc-650c-4c38-b35d-4f730a135f76","order_by":0,"name":"Nan 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