Aptamer-based fluorometric determination of norovirus using a paper-based microfluidic device
- 761 Downloads
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.
KeywordsBiosensor Aptamer Norovirus Paper-based microfluidic device Nitrocellulose membrane Multi-walled carbon nanotubes Graphene oxide
enzyme-linked immunosorbent assay
fluorescence resonance energy transfer
limit of detection
multi-walled carbon nanotubes
reverse transcription polymerase chain reaction
The authors sincerely thank the Natural Sciences and Engineering Research Council of Canada (400705) for funding this study.
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.
Compliance with ethical standards
The authors declare that they have no competing interests.
- 2.Koo HL, Ajami N, Atmar RL, DuPont HL (2010) Noroviruses: the principal cause of foodborne disease worldwide. Discov Med 10(50):61Google Scholar
- 4.Jiang XI, Wang MI, Graham DY, Estes MK (1992) Expression, self-assembly, and antigenicity of the Norwalk virus capsid protein. J Virol 66(11):6527–6532Google Scholar
- 5.Hayashi YU, Ando TA, Utagawa ET, Sekine SE, Okada SH, Yabuuchi K, Miki T, Ohashi M (1989) Western blot (immunoblot) assay of small, round-structured virus associated with an acute gastroenteritis outbreak in Tokyo. J Clin Microbiol 27(8):1728–1733Google Scholar
- 6.Vinjé J, Vennema H, Maunula L, von Bonsdorff CH, Hoehne M, Schreier E, Richards A, Green J, Brown D, Beard SS, Monroe SS (2003) International collaborative study to compare reverse transcriptase PCR assays for detection and genotyping of noroviruses. J Clin Microbiol 41(4):1423–1433CrossRefGoogle Scholar
- 29.Thomsen V, Schatzlein D, Mercuro D (2003) Limits of detection in spectroscopy. Spectroscopy 18(12):112–114Google Scholar
- 30.Neethirajan S, Ahmed SR, Chand R, Buozis J, Nagy É (2017) Recent advances in biosensor development for foodborne virus detection. Nano 1:272–295Google Scholar
- 32.Todd EC (2016) Foodborne diseases: case Studies of outbreaks in the agri-food industries. CRC Press, (318–240)Google Scholar