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Portable bead-based fluorescence detection system for multiplex nucleic acid testing: a case study with Bacillus anthracis

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Abstract

This paper describes the design, functioning and use of a portable detection platform for multiplex nucleic acid testing. The system features a bead-supported DNA hybridization assay performed inside a microfluidic cartridge. Polystyrene particles modified with DNA capture probes are confined in the detection area and exposed to a solution of fluorescently labeled target DNA strands. The cartridge, fabricated from inexpensive thermoplastic polymers, allows for conducting up to eight assays in parallel. The detection instrument is equipped with a pneumatic module and a manifold lid serving as an interface to mediate fluid displacement on the cartridge. The fluorescence signal deriving from each assay is recorded by a semi-confocal fluorescence reader embedded in the detection platform. The compact design of the instrument and its level of integration make it possible to obtain an analytical result in less than 15 min, while only few manual steps need to be performed in between. A proof-of-concept demonstration involving Cy3-labeled, PCR-amplified genomic DNA confirms the ability to detect Bacillus anthracis in a multiplexed single-assay format using lef and capC genes. Limits of quantification are on the order of 1 × 109 copies/μL for lef targets.

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Acknowledgments

This work was supported in part by Canada’s Chemical, Biological, Radiological, and Nuclear Research and Technology Initiative (CRTI) under the program Portable Biological Agent Detection System (06-0187TD). We would like to thank our colleagues and collaborators for useful discussions and technical assistance: Kien-Mun Lau, Maxence Mounier, François Normandin, Boris Le Drogoff, Jean-Guy Allard, Emmanuel Roy, and Hélène Roberge from CNRC Boucherville; Arnold Kell, Chantal Paquet, and Benoît Simard from the Steacie Institute for Molecular Sciences (CNRC, Ottawa, ON); Thompson Tang, William Lee, and Doug Bader from Defense Research and Development Canada (Suffield, Medicine Hat, AB); Ian Summerell from the Royal Canadian Mounted Police (Orleans, ON), Louis Bryden and Michael Mulvey from the Public Health Agency of Canada (Winnipeg, MB); and David Béliveau-Viel, Sébastien Dubus, Even Lemieux, and Prof. Mario Leclerc from Université Laval. We are grateful to Caroline Vachon and Michel M. Dumoulin from CNRC Boucherville for their support.

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Authors and Affiliations

  1. Département de chimie et centre d’optique, photonique et laser (COPL), Université Laval, Quebec, QC, G1V 0A6, Canada

    Jean-François Gravel, Sébastien Chapdelaine, Hugo-Pierre Poirier-Richard, Alexandre Grégoire & Denis Boudreau

  2. Conseil national de recherches Canada (CNRC), Boucherville, QC, J4B 6Y4, Canada

    Matthias Geissler, Benoît Voisin & Teodor Veres

  3. Centre de recherche en infectiologie de l’Université Laval, Centre de recherche du CHUQ, Quebec, QC, G1V 4G2, Canada

    Karel Boissinot, Isabelle Charlebois, Maurice Boissinot & Michel G. Bergeron

Authors
  1. Jean-François Gravel
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  2. Matthias Geissler
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  3. Sébastien Chapdelaine
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  4. Karel Boissinot
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  5. Benoît Voisin
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  6. Isabelle Charlebois
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  7. Hugo-Pierre Poirier-Richard
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  8. Alexandre Grégoire
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  9. Maurice Boissinot
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  10. Michel G. Bergeron
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  11. Teodor Veres
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  12. Denis Boudreau
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Corresponding authors

Correspondence to Teodor Veres or Denis Boudreau.

Additional information

Jean-François Gravel and Matthias Geissler have contributed equally to this work.

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Gravel, JF., Geissler, M., Chapdelaine, S. et al. Portable bead-based fluorescence detection system for multiplex nucleic acid testing: a case study with Bacillus anthracis . Microfluid Nanofluid 16, 1075–1087 (2014). https://doi.org/10.1007/s10404-013-1273-y

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  • DOI: https://doi.org/10.1007/s10404-013-1273-y

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