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dc.contributor.authorHuang, Shichuen_US
dc.contributor.authorDo, Jaephilen_US
dc.contributor.authorMahalanabis, Madhumitaen_US
dc.contributor.authorFan, Andyen_US
dc.contributor.authorZhao, Leien_US
dc.contributor.authorJepeal, Lisaen_US
dc.contributor.authorSingh, Satish K.en_US
dc.contributor.authorKlapperich, Catherine M.en_US
dc.date.accessioned2020-05-11T15:13:50Z
dc.date.available2020-05-11T15:13:50Z
dc.date.issued2013-03-28
dc.identifierhttp://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000317262200075&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=6e74115fe3da270499c3d65c9b17d654
dc.identifier.citationShichu Huang, Jaephil Do, Madhumita Mahalanabis, Andy Fan, Lei Zhao, Lisa Jepeal, Satish K. Singh, Catherine M. Klapperich. 2013. "Low Cost Extraction and Isothermal Amplification of DNA for Infectious Diarrhea Diagnosis." PLOS ONE, Volume 8, Issue 3. https://doi.org/10.1371/journal.pone.0060059
dc.identifier.issn1932-6203
dc.identifier.urihttps://hdl.handle.net/2144/40736
dc.description.abstractIn order to counter the common perception that molecular diagnostics are too complicated to work in low resource settings, we have performed a difficult sample preparation and DNA amplification protocol using instrumentation designed to be operated without wall or battery power. In this work we have combined a nearly electricity-free nucleic acid extraction process with an electricity-free isothermal amplification assay to detect the presence of Clostridium difficile (C. difficile) DNA in the stool of infected patients. We used helicase-dependent isothermal amplification (HDA) to amplify the DNA in a low-cost, thermoplastic reaction chip heated with a pair of commercially available toe warmers, while using a simple Styrofoam insulator. DNA was extracted from known positive and negative stool samples. The DNA extraction protocol utilized an air pressure driven solid phase extraction device run using a standard bicycle pump. The simple heater setup required no electricity or battery and was capable of maintaining the temperature at 65°C±2°C for 55 min, suitable for repeatable HDA amplification. Experiments were performed to explore the adaptability of the system for use in a range of ambient conditions. When compared to a traditional centrifuge extraction protocol and a laboratory thermocycler, this disposable, no power platform achieved approximately the same lower limit of detection (1.25×10−2 pg of C. difficile DNA) while requiring much less raw material and a fraction of the lab infrastructure and cost. This proof of concept study could greatly impact the accessibility of molecular assays for applications in global health.en_US
dc.description.sponsorshipThis work was funded by NIH Grant R21 AI71261 to CMK and SS. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. (R21 AI71261 - NIH)en_US
dc.format.extent10 pagesen_US
dc.languageEnglish
dc.language.isoen_US
dc.publisherPublic Library of Scienceen_US
dc.relation.ispartofPLOS ONE
dc.rights© 2013 Huang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en_US
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectScience & technologyen_US
dc.subjectMultidisciplinary sciencesen_US
dc.subjectImmunochromatographic strip testen_US
dc.subjectDisposable detection deviceen_US
dc.subjectReal-time PCRen_US
dc.subjectOn-a-chipen_US
dc.subjectClostridium difficleen_US
dc.subjectGlobal healthen_US
dc.subjectIdentificationen_US
dc.subjectMicrofluidicsen_US
dc.subjectTechnologiesen_US
dc.subjectSystemsen_US
dc.subjectDNAen_US
dc.subjectDiarrheaen_US
dc.subjectHumansen_US
dc.subjectNucleic acid amplification techniquesen_US
dc.subjectTemperatureen_US
dc.titleLow cost extraction and isothermal amplification of DNA for infectious diarrhea diagnosisen_US
dc.typeArticleen_US
dc.description.versionFinal published versionen_US
dc.identifier.doi10.1371/journal.pone.0060059
pubs.elements-sourceweb-of-scienceen_US
pubs.notesEmbargo: Not knownen_US
pubs.organisational-groupBoston Universityen_US
pubs.organisational-groupBoston University, College of Engineeringen_US
pubs.organisational-groupBoston University, College of Engineering, Department of Biomedical Engineeringen_US
pubs.organisational-groupBoston University, School of Medicineen_US
pubs.publication-statusPublisheden_US
dc.identifier.orcid0000-0001-6103-849X (Klapperich, Catherine M)
dc.identifier.mycv34095


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© 2013 Huang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Except where otherwise noted, this item's license is described as © 2013 Huang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.