Infection on a chip: a microscale platform for simple and sensitive cell-based virus assays

Biomed Microdevices. 2009 Jun;11(3):565-70. doi: 10.1007/s10544-008-9263-7.

Abstract

The plaque assay has long served as the "gold standard" to measure virus infectivity and test antiviral drugs, but the assay is labor-intensive, lacks sensitivity, uses excessive reagents, and is hard to automate. Recent modification of the assay to exploit flow-enhanced virus spread with quantitative imaging has increased its sensitivity. Here we performed flow-enhanced infection assays in microscale channels, employing passive fluid pumping to inoculate cell monolayers with virus and drive infection spread. Our test of an antiviral drug (5-fluorouracil) against vesicular stomatitis virus infections of BHK cell monolayers yielded a two-fold improvement in sensitivity, relative to the standard assay based on plaque counting. The reduction in scale, simplified fluid handling, image-based quantification, and higher assay sensitivity will enable infection measurements for high-throughput drug screening, sero-conversion testing, and patient-specific diagnosis of viral infections.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiviral Agents / pharmacology
  • Cell Culture Techniques
  • Cell Line
  • Cricetinae
  • Dose-Response Relationship, Drug
  • Equipment Design
  • Fluorescent Dyes / metabolism
  • Fluorouracil / pharmacology
  • Immunohistochemistry
  • Indoles / metabolism
  • Microfluidic Analytical Techniques / instrumentation*
  • Microfluidic Analytical Techniques / methods
  • Microtechnology
  • Rhabdoviridae Infections / drug therapy*
  • Rhabdoviridae Infections / immunology
  • Rhabdoviridae Infections / virology
  • Sensitivity and Specificity
  • Vesicular stomatitis Indiana virus / drug effects
  • Vesicular stomatitis Indiana virus / growth & development*
  • Vesicular stomatitis Indiana virus / immunology*
  • Viral Plaque Assay / standards*

Substances

  • Antiviral Agents
  • Fluorescent Dyes
  • Indoles
  • DAPI
  • Fluorouracil