Development of a system for on-disc isothermal in vitro amplification and detection of bacterial RNA

Des Brennan, Helena Coughla, Eoin Clancy, Nikolay Dimov, Thomas Barry, David Kinahan, Jens Ducree, Terry J. Smith, Paul Galvin

Research output: Chapter in Book/Report/Conference proceedingConference contribution

1 Citation (Scopus)

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. The motivation for such systems is to implement fast point of care tests in a healthcare setting. Rapid H. Influenzae identification is crucial for efficient treatment to avoid serious health effects (e.g. childhood meningitis). Molecular diagnostic tests offer the sensitivity and speed required for such applications.

Original languageEnglish
Title of host publication2017 IEEE Biomedical Circuits and Systems Conference, BioCAS 2017 - Proceedings
PublisherInstitute of Electrical and Electronics Engineers (IEEE)
Pages1-4
Number of pages4
ISBN (Electronic)9781509058037
DOIs
Publication statusPublished - 23 Mar 2018
Event2017 IEEE Biomedical Circuits and Systems Conference, BioCAS 2017 - Torino, Italy
Duration: 19 Oct 201721 Oct 2017

Publication series

Name2017 IEEE Biomedical Circuits and Systems Conference, BioCAS 2017 - Proceedings
Volume2018-January

Conference

Conference2017 IEEE Biomedical Circuits and Systems Conference, BioCAS 2017
Country/TerritoryItaly
CityTorino
Period19/10/1721/10/17

Keywords

  • centrifugal disc
  • fluorescence detection
  • IR heating
  • Isothermal amplification
  • tmRNA

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