Development of an on-disc isothermal in vitro amplification and detection of bacterial RNA

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dc.contributor.authorBrennan, Des
dc.contributor.authorCoughlan, Helena
dc.contributor.authorClancy, Eoin
dc.contributor.authorDimov, Nikolay
dc.contributor.authorBarry, Thomas
dc.contributor.authorKinahan, David
dc.contributor.authorDucrée, Jens
dc.contributor.authorSmith, Terry J.
dc.contributor.authorGalvin, Paul
dc.contributor.funderScience Foundation Irelanden
dc.date.accessioned2023-03-16T15:43:14Z
dc.date.available2023-03-16T15:43:14Z
dc.date.issued2016-08-07
dc.date.updated2023-03-16T15:34:29Z
dc.description.abstractWe present a centrifugal microfluidic "Lab-on-a-Disc" (LoaD) system capable of implementing nucleic acid in vitro amplification using non-contact heating and fluorescence detection. The system functionality is verified by implementing a Nucleic Acid Sequence Based Amplification (NASBA) reaction, targeting the tmRNA transcript of Haemophilus influenzae. The NASBA assay incorporates fluorescent molecular beacon probes reporting target tmRNA amplification for endpoint detection. The system implements non contact IR heating to heat the NASBA reaction to the required target temperatures during denaturation and amplification steps. The LoaD control system facilitates spin speed and chamber positioning for heating and fluorescence detection. The LoaD alignment system uses magnetic fields to locate and lock the chamber in the required position (heating or detection). The NASBA assay was implemented on the system using Haemophilus influenzae tmRNA over the range 10(2)-10(4) cell equivalent (CE) units. For comparison, identical qNASBA assays were implemented on a Roche LightCycler 2.0 over this concentration range.en
dc.description.statusPeer revieweden
dc.description.versionAccepted Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationBrennan, D., Coughlan, H., Clancy, E., Dimov, N., Barry, T., Kinahan, D., Ducrée, J., Smith, T. J. and Galvin, P. (2016) 'Development of an on-disc isothermal in vitro amplification and detection of bacterial RNA', Sensors and Actuators, B: Chemical, 239, pp. 235-242. doi: 10.1016/j.snb.2016.08.018en
dc.identifier.doi10.1016/j.snb.2016.08.018en
dc.identifier.endpage242en
dc.identifier.issn0925-4005
dc.identifier.journaltitleSensors and Actuators, B: Chemicalen
dc.identifier.startpage235en
dc.identifier.urihttps://hdl.handle.net/10468/14322
dc.identifier.volume239en
dc.language.isoenen
dc.publisherElsevier B.V.en
dc.relation.projectinfo:eu-repo/grantAgreement/SFI/SFI Centre for Science Engineering and Technology (CSET)/10/CE/B1821/IE/CSET BDI: Biomedical Diagnostics Institute/en
dc.rights© 2016, Elsevier B.V. All rights reserved. This manuscript version is made available under the CC BY-NC-ND 4.0 license.en
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/en
dc.subjectIsothermal amplificationen
dc.subjectLab-on-a-Disc(LoaD)en
dc.subjecttmRNAen
dc.subjectIR heatingen
dc.subjectFluorescence detectionen
dc.titleDevelopment of an on-disc isothermal in vitro amplification and detection of bacterial RNAen
dc.typeArticle (peer-reviewed)en
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