University of Hertfordshire

Documents

  • Des Brennan
  • Helena Coughlan
  • Eoin Clancy
  • Nikolay Dimov
  • Thomas Barry
  • David Kinahan
  • Jens Ducrée
  • Terry J. Smith
  • Paul Galvin
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Original languageEnglish
Number of pages8
Pages (from-to)235-242
JournalSensors and Actuators, B: Chemical
Journal publication date1 Feb 2017
Volume239
Early online date3 Aug 2016
DOIs
Publication statusPublished - 1 Feb 2017

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.

Notes

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.

ID: 13219697