Portable Detection of Monkeypox virus Clades using Bridging LAMP

Portable Detection of Monkeypox virus Clades using Bridging LAMP | 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 Portable Detection of Monkeypox virus Clades using Bridging LAMP Karen Anderson, Mark Knappenberger, Siril Arockiam, Vi Nguyen, and 12 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9014850/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 Orthopoxvirus monkeypox (MPXV) is a zoonotic pathogen with two major clades that cause outbreaks in humans. Differentiation between the historically more clinically severe clade I and clade IIb is needed for effective public health response, such as in the ongoing 2022-2025 MPXV multi-country outbreaks. We have developed three loop-mediated isothermal amplification (LAMP) assays that differentiate between clade I, clade IIa/IIb MPXV, and broader Orthopoxvirus species by exploiting viral genomic rearrangements and amplicon length limitations inherent to LAMP. We demonstrate clade specificity between clade I and clade II viral cell culture lysates and rapid detection of crude lesion specimens isolated from 45 patients during the latest MPXV USA 2022-2025 outbreaks. MPXV genomic DNA can be detected in as little as 10 minutes and clade-specific results detected in under an hour with limited instrumentation. We adapt a portable microfluidic system for point-of-need outbreak responses to zoonotic orthopoxvirus. Health sciences/Biomarkers/Diagnostic markers Biological sciences/Biological techniques/Sensors and probes/Oligonucleotide probes Full Text Additional Declarations Yes there is potential Competing Interest. K.S.A. and J.B.C. are founding members of Flex Biotech. M.D.K., and K.S.A. have a patent application for detection of Orthopoxvirus monkeypox using the assays described in this manuscript. The authors have no other conflicts of interest to disclose. The findings and conclusions in this report are those of the authors and do not necessarily represent the official position of the Centers for Disease Control and Prevention. 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-9014850","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":638889222,"identity":"7e44f8dd-511c-47f6-b224-29ef95042b9f","order_by":0,"name":"Karen Anderson","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/0lEQVRIie3NMUvDQBTA8RcOzuVp1nec+hkCB0Fo0a+SQ0iWIoJLQcFMcdKuzbfo6Bgo9JaIazYphXNxSOkoiCkBnc7q5nD/5d3wfu8AfL5/GH29qn5g2E/2e3Io8r+SYVTtIOLuabm6hNOLA3O/atePgMoYSzAe6NxBJGZKTeH8StQmFmUNGNejmKDOnOQYUi4RmJ41KWf7RUeqEaegmLtJaPfeEW717MWyzZaoyaul4MNNJHXHEebdLxzklkSUxBTkbiKmlkmMjC7rlIuyIKTmTZ0ki0y5CD2nwQbH1/rBLFi7LoZn4SRbNu3N4MhF+qLvE/1Iflz3+Xw+364+AXd5T67/qOkZAAAAAElFTkSuQmCC","orcid":"","institution":"Virginia G. 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