Norovirus detection in water samples at the level of single virus copies per microliter using a smartphone-based fluorescence microscope
Breshears, Lane E.
Jennings, Christian M.
Morrison, Christina M.
Betancourt, Walter Q.
Reynolds, Kelly A.
AffiliationDepartment of Biosystems Engineering, The University of Arizona
Department of Biomedical Engineering, The University of Arizona
Department of Environmental Science, The University of Arizona
Mel and Enid Zuckerman College of Public Health, The University of Arizona
MetadataShow full item record
CitationChung, S., Breshears, L.E., Gonzales, A. et al. Norovirus detection in water samples at the level of single virus copies per microliter using a smartphone-based fluorescence microscope. Nat Protoc (2021). https://doi.org/10.1038/s41596-020-00460-7
RightsCopyright © 2021, The Author(s), under exclusive licence to Springer Nature Limited.
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AbstractNorovirus is a widespread public health threat and has a very low infectious dose. This protocol presents the extremely sensitive mobile detection of norovirus from water samples using a custom-built smartphone-based fluorescence microscope and a paper microfluidic chip. Antibody-conjugated fluorescent particles are immunoagglutinated and spread over the paper microfluidic chip by capillary action for individual counting using a smartphone-based fluorescence microscope. Smartphone images are analyzed using intensity- and size-based thresholding for the elimination of background noise and autofluorescence as well as for the isolation of immunoagglutinated particles. The resulting pixel counts of particles are correlated with the norovirus concentration of the tested sample. This protocol provides detailed guidelines for the construction and optimization of the smartphone- and paper-based assay. In addition, a 3D-printed enclosure is presented to incorporate all components in a dark environment. On-chip concentration and the assay of higher concentrations are presented to further broaden the assay range. This method is the first to be presented as a highly sensitive mobile platform for norovirus detection using low-cost materials. With all materials and reagents prepared, a single standard assay takes under 20 min. Although the method described is used for detection of norovirus, the same protocol could be adapted for detection of other pathogens by using different antibodies. © 2021, The Author(s), under exclusive licence to Springer Nature Limited.
Note6 month embargo; published 29 January 2021
VersionFinal accepted manuscript
SponsorsNational Science Foundation