Rapid Detection of Norovirus Using Paper-based Microfluidic Device

Noroviruses (NoV) are the leading cause of outbreak of acute gastroenteritis worldwide. A substantial effort has been made in the development of analytical devices for rapid and sensitive food safety monitoring via the detection of foodborne bacteria, viruses and parasites. Conventional analytical approaches for noroviruses suffer from some critical weaknesses: labor-intensive, time-consuming, and relatively low sensitivity. In this study, we developed a rapid and highly sensitive biosensor towards point-of-care device for noroviruses based on 6-carboxyfluorescein (6-FAM) labeled aptamer and nanomaterials, multi-walled carbon nanotubes (MWCNTs) and graphene oxide (GO). In an assay, the fluorescence of 6-FAM labeled aptamer was quenched by MWCNTs or GO via fluorescence resonance energy transfer (FRET). In the presence of norovirus, the fluorescence would be recovered due to the release of the 6-FAM labeled aptamer from MWCNTs or GO. An easy-to-make paper-based microfluidic platform made by nitrocellulose membrane was used to conduct the assay. The quantitative detection of norovirus virus-like particles (NoV VLPs) was successfully performed. A linear range of 0-12.9 g/mL with a detection limit of 40 pM and 30 pM was achieved for the MWCNTs and GO based paper sensors, respectively. The results suggested the developed paper-based microfluidic device is simple, cost-effective and holds the potential of rapid in situ visual determination for noroviruses with remarkable sensitivity and specificity, which provides a new way for early identification of NoV and thereby an early intervention for preventing the spread of an outbreak.