Infection on a chip: a microscale platform for simple and sensitive cell-based virus assays.
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.
Zhu
Ying
Y
Department of Chemical and Biological Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.
Warrick
Jay W
JW
Haubert
Kathryn
K
Beebe
David J
DJ
Yin
John
J
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D018929:Cell Culture Techniques; D002460:Cell Line; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D004867:Equipment Design; D005456:Fluorescent Dyes / Q000378:metabolism; D005472:Fluorouracil / Q000494:pharmacology; D007150:Immunohistochemistry; D007211:Indoles / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D055616:Microtechnology; D018353:Rhabdoviridae Infections / Q000188:drug therapy* / Q000276:immunology / Q000821:virology; D012680:Sensitivity and Specificity; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects / Q000254:growth & development* / Q000276:immunology*; D010948:Viral Plaque Assay / Q000592:standards*
D000998:Antiviral Agents; D005456:Fluorescent Dyes; D007211:Indoles; C007293:DAPI; D005472:Fluorouracil