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Detection of Respiratory Viruses with Plastic High Throughput Screening Devices

Published online by Cambridge University Press:  01 February 2011

Zhengshan Zhao
Affiliation:
National Research Council of Canada, Industrial Materials Institute, 75 de Mortagne, Boucherville, J4B 6Y4, Canada, (450)-641-5023, (450)-641-5105
Gerardo A. Diaz-Quijada
Affiliation:
[email protected], National Research Council of Canada, Industrial Materials Institute, 75 de Mortagne, Boucherville, J4B 6Y4, Canada
Régis Peytavi
Affiliation:
[email protected], Université Laval, Centre de recherche en infectiologie, 2705 Boul. Laurier, Sainte Foy, G1V 4G2, Canada
Éric LeBlanc
Affiliation:
[email protected], Université Laval, Centre de recherche en infectiologie, 2705 Boul. Laurier, Sainte Foy, G1V 4G2, Canada
Johanne Frenette
Affiliation:
[email protected], Université Laval, Centre de recherche en infectiologie, 2705 Boul. Laurier, Sainte Foy, G1V 4G2, Canada
Guy Boivin
Affiliation:
[email protected], Université Laval, Centre de recherche en infectiologie, 2705 Boul. Laurier, Sainte Foy, G1V 4G2, Canada
Jim V. Zoval
Affiliation:
[email protected], University of California, Department of Mechanical and Aerospace Engineering, Irvine, CA, 92697-3975, United States
Marc Madou
Affiliation:
[email protected], University of California, Department of Mechanical and Aerospace Engineering, Irvine, CA, 92697-3975, United States
Michel M. Dumoulin
Affiliation:
[email protected], National Research Council of Canada, Industrial Materials Institute, 75 de Mortagne, Boucherville, J4B 6Y4, Canada
Teodor Veres
Affiliation:
[email protected], National Research Council of Canada, Industrial Materials Institute, 75 de Mortagne, Boucherville, J4B 6Y4, Canada
Michel G. Bergeron
Affiliation:
[email protected], Université Laval, Centre de recherche en infectiologie, 2705 Boul. Laurier, Sainte Foy, G1V 4G2, Canada
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Abstract

Microarrays have become one of the most convenient tools for high throughput screening and have catalyzed major advances in genomics and proteomics. Other important applications can be found in medical diagnostics, detection of biothreats, drug discovery, etc. Integration of microarrays with microfluidic devices can be highly advantageous in terms of portability, shorter analysis time and lower consumption of expensive biological analytes. Since fabrication of microfluidic devices using traditional materials such as glass is rather expensive, there is a high interest in employing polymeric materials as a low cost alternative suitable for mass production. We present proof-of-concept DNA arrays on a plastic platform for the detection of four important respiratory pathogens: Influenza A virus, respiratory syncytial virus, human enterovirus, and human metapneumovirus.

This was accomplished by amplifying the genetic material from the viruses and simultaneously labeling the amplicons with a fluorescent dye (Cy3) via a highly sensitive multiplex Reverse Transcription Polymerase Chain Reaction (RT-PCR). The resultant RT-PCR product was hybridized, without further purification, with an array of specific oligonucleotide probes (20 mers) that had been covalently bound to a plastic substrate. Results indicate a high signal to background ratio that is comparable to commercially available microarray glass slides. In addition, 5 minute hybridization on this plastic substrate has been demonstrated using a centrifugal microfluidic platform, paving the way to a rapid medical diagnostic device for point-of-care use that is based on a low-cost portable Micro-Total-Analysis-System (μ-TAS).

Type
Research Article
Copyright
Copyright © Materials Research Society 2007

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References

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