University of Hertfordshire

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

Research output: Contribution to journalArticlepeer-review

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Development of an on-disc isothermal in vitro amplification and detection of bacterial RNA. / Brennan, Des; Coughlan, Helena; Clancy, Eoin; Dimov, Nikolay; Barry, Thomas; Kinahan, David; Ducrée, Jens; Smith, Terry J.; Galvin, Paul.

In: Sensors and Actuators, B: Chemical, Vol. 239, 01.02.2017, p. 235-242.

Research output: Contribution to journalArticlepeer-review

Harvard

Brennan, D, Coughlan, H, Clancy, E, Dimov, N, Barry, T, Kinahan, D, Ducrée, J, Smith, TJ & Galvin, P 2017, 'Development of an on-disc isothermal in vitro amplification and detection of bacterial RNA', Sensors and Actuators, B: Chemical, vol. 239, pp. 235-242. https://doi.org/10.1016/j.snb.2016.08.018

APA

Brennan, D., Coughlan, H., Clancy, E., Dimov, N., Barry, T., Kinahan, D., Ducrée, J., Smith, T. J., & Galvin, P. (2017). Development of an on-disc isothermal in vitro amplification and detection of bacterial RNA. Sensors and Actuators, B: Chemical, 239, 235-242. https://doi.org/10.1016/j.snb.2016.08.018

Vancouver

Author

Brennan, Des ; Coughlan, Helena ; Clancy, Eoin ; Dimov, Nikolay ; Barry, Thomas ; Kinahan, David ; Ducrée, Jens ; Smith, Terry J. ; Galvin, Paul. / Development of an on-disc isothermal in vitro amplification and detection of bacterial RNA. In: Sensors and Actuators, B: Chemical. 2017 ; Vol. 239. pp. 235-242.

Bibtex

@article{e8ed62b32288428eb979391b6a1a0873,
title = "Development of an on-disc isothermal in vitro amplification and detection of bacterial RNA",
abstract = "We 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 102–104 cell equivalent (CE) units. For comparison, identical qNASBA assays were implemented on a Roche LightCycler 2.0 over this concentration range.",
keywords = "Fluorescence detection, IR heating, Isothermal amplification, Lab-on-a-Disc(LoaD), tmRNA",
author = "Des Brennan and Helena Coughlan and Eoin Clancy and Nikolay Dimov and Thomas Barry and David Kinahan and Jens Ducr{\'e}e and Smith, {Terry J.} and Paul Galvin",
note = "This document is the Accepted Manuscript version, made available under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License CC BY NC-ND 4.0 (http://creativecommons.org/licenses/by-nc-nd/4.0/). The final, published version is available online at doi: https://doi.org/10.1016/j.snb.2016.08.018. Published by Elsevier B. V. ",
year = "2017",
month = feb,
day = "1",
doi = "10.1016/j.snb.2016.08.018",
language = "English",
volume = "239",
pages = "235--242",
journal = "Sensors and Actuators B: Chemical",
issn = "0925-4005",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Development of an on-disc isothermal in vitro amplification and detection of bacterial RNA

AU - Brennan, Des

AU - Coughlan, Helena

AU - Clancy, Eoin

AU - Dimov, Nikolay

AU - Barry, Thomas

AU - Kinahan, David

AU - Ducrée, Jens

AU - Smith, Terry J.

AU - Galvin, Paul

N1 - This document is the Accepted Manuscript version, made available under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License CC BY NC-ND 4.0 (http://creativecommons.org/licenses/by-nc-nd/4.0/). The final, published version is available online at doi: https://doi.org/10.1016/j.snb.2016.08.018. Published by Elsevier B. V.

PY - 2017/2/1

Y1 - 2017/2/1

N2 - We 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 102–104 cell equivalent (CE) units. For comparison, identical qNASBA assays were implemented on a Roche LightCycler 2.0 over this concentration range.

AB - We 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 102–104 cell equivalent (CE) units. For comparison, identical qNASBA assays were implemented on a Roche LightCycler 2.0 over this concentration range.

KW - Fluorescence detection

KW - IR heating

KW - Isothermal amplification

KW - Lab-on-a-Disc(LoaD)

KW - tmRNA

UR - http://www.scopus.com/inward/record.url?scp=84982854033&partnerID=8YFLogxK

U2 - 10.1016/j.snb.2016.08.018

DO - 10.1016/j.snb.2016.08.018

M3 - Article

AN - SCOPUS:84982854033

VL - 239

SP - 235

EP - 242

JO - Sensors and Actuators B: Chemical

JF - Sensors and Actuators B: Chemical

SN - 0925-4005

ER -