Aptamer-based fluorometric determination of norovirus using a paper-based microfluidic device

Abstract

The authors describe a rapid and highly sensitive point-of-care device for rapid determination of noroviruses, a leading cause of acute gastroenteritis. The assay is based on the use of a norovirus-specific aptamer labeled with 6-carboxyfluorescein, and of multi-walled carbon nanotubes (MWCNT) and graphene oxide (GO). The fluorescence of the 6-FAM labeled aptamer is quenched by MWCNT or GO. In the presence of norovirus, fluorescence is recovered due to the release of the labeled aptamer from MWCNT or GO. An easy-to-make paper-based microfluidic platform was developed using a nitrocellulose membrane. The quantitation of norovirus was successfully performed. The linear range extends from 13 ng·mL−1 to 13 μg·mL−1 of norovirus. The detection limits are 4.4 ng·mL−1 and 3.3 ng·mL−1, respectively, when using MWCNT or GO. The device is simple and cost-effective, and holds the potential of rapid in-situ visual determination of noroviruses with remarkable sensitivity and specificity. Hence, it provides a new method for early identification of norovirus and a tool for early intervention when preventing the spread of an outbreak.

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Abbreviations

6-FAM:

6-carboxyfluorescein

ELISA:

enzyme-linked immunosorbent assay

FRET:

fluorescence resonance energy transfer

GO:

graphene oxide

LoD:

limit of detection

LFAs:

lateral-flow assays

MWCNT:

multi-walled carbon nanotubes

POCT:

point-of-care testing

RT-PCR:

reverse transcription polymerase chain reaction

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Acknowledgements

The authors sincerely thank the Natural Sciences and Engineering Research Council of Canada (400705) for funding this study.

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Contributions

XW and SN designed the study; XW performed experiments, acquired and analyzed data, XW and SN drafted and edited the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Suresh Neethirajan.

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The authors declare that they have no competing interests.

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Weng, X., Neethirajan, S. Aptamer-based fluorometric determination of norovirus using a paper-based microfluidic device. Microchim Acta 184, 4545–4552 (2017). https://doi.org/10.1007/s00604-017-2467-x

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Keywords

  • Biosensor
  • Aptamer
  • Norovirus
  • Paper-based microfluidic device
  • Nitrocellulose membrane
  • Multi-walled carbon nanotubes
  • Graphene oxide