Issue 19, 2015

Rapid, targeted and culture-free viral infectivity assay in drop-based microfluidics

Abstract

A key viral property is infectivity, and its accurate measurement is crucial for the understanding of viral evolution, disease and treatment. Currently viral infectivity is measured using plaque assays, which involve prolonged culturing of host cells, and whose measurement is unable to differentiate between specific strains and is prone to low number fluctuation. We developed a rapid, targeted and culture-free infectivity assay using high-throughput drop-based microfluidics. Single infectious viruses are incubated in a large number of picoliter drops with host cells for one viral replication cycle followed by in-drop gene-specific amplification to detect infection events. Using murine noroviruses (MNV) as a model system, we measure their infectivity and determine the efficacy of a neutralizing antibody for different variants of MNV. Our results are comparable to traditional plaque-based assays and plaque reduction neutralization tests. However, the fast, low-cost, highly accurate genomic-based assay promises to be a superior method for drug screening and isolation of resistant viral strains. Moreover our technique can be adapted to measuring the infectivity of other pathogens, such as bacteria and fungi.

Graphical abstract: Rapid, targeted and culture-free viral infectivity assay in drop-based microfluidics

Supplementary files

Article information

Article type
Paper
Submitted
17 May 2015
Accepted
14 Aug 2015
First published
17 Aug 2015

Lab Chip, 2015,15, 3934-3940

Rapid, targeted and culture-free viral infectivity assay in drop-based microfluidics

Y. Tao, A. Rotem, H. Zhang, C. B. Chang, A. Basu, A. O. Kolawole, S. A. Koehler, Y. Ren, J. S. Lin, J. M. Pipas, A. B. Feldman, C. E. Wobus and D. A. Weitz, Lab Chip, 2015, 15, 3934 DOI: 10.1039/C5LC00556F

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