Human enteric viruses can be highly infectious and thus capable of causing disease upon ingestion of low doses ranging from 10<sup>0</sup> to 10<sup>2</sup> virions. Norovirus is a good example with a minimum infectious dose as low as a few tens of virus particles, i.e. below femtogram scale. Norovirus detection from commonly implicated environmental matrices (water and food) involves complicated concentration of viruses and/or amplification of its genetic materials, thus rendering detection approaches not feasible for field applications. In this work, norovirus detection was performed on a microfluidic paper analytic device (μPAD) without using any sample concentration or nucleic acid amplification steps, by directly imaging and counting on-paper aggregation of antibody-conjugated, fluorescent submicron particles. An in-house developed smartphone-based fluorescence microscope and an image processing algorithm isolated only the particles aggregated by antibody-antigen binding, leading to an extremely low limit of norovirus detection, as low as 1 genome copy/μL in deionized water and 10 genome copies/μL in reclaimed wastewater.

Smartphone-base paper microfluidic particulometry of norovirus from environmental water samples at single copy level

2019 Fall National Meeting

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