tetano
Editor, Senior Moderator
J Virol Methods. 2011 Dec 3. [Epub ahead of print]
A quantitative comet infection assay for influenza virus.
Lindsay SM, Timm A, Yin J.
Source
Department of Chemical and Biological Engineering, Systems Biology Theme, Wisconsin Institute for Discovery, University of Wisconsin - Madison, 330N Orchard Street, Madison, WI 53715, United States.
Abstract
The virus comet assay is a cell-based virulence assay used to evaluate an antiviral drug or antibody against a target virus. The comet assay differs from the plaque assay in allowing spontaneous flows in 6-well plates to spread virus. When implemented quantitatively the comet assay has been shown to have an order-of-magnitude greater sensitivity to antivirals than the plaque assay. In this study, a quantitative comet assay for influenza virus is demonstrated, and is shown to have a 13-fold increase in sensitivity to ribavirin. AX4 cells (MDCK cells with increased surface concentration of α2-6 sialic acid, the influenza virus receptor) have reduced the comet size variability relative to MDCK cells, making them a better host cell for use in this assay. Because of enhanced antiviral sensitivity in flow-based assays, less drug is required, which could lead to lower reagent costs, reduced cytotoxicity, and fewer false-negative drug screen results. The comet assay also serves as a readout of flow conditions in the well. Observations from comets formed at varying humidity levels indicate a role for evaporation in the mechanism of spontaneous fluid flow in wells.
Copyright ? 2011. Published by Elsevier B.V.
PMID:
22155578
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22155578
A quantitative comet infection assay for influenza virus.
Lindsay SM, Timm A, Yin J.
Source
Department of Chemical and Biological Engineering, Systems Biology Theme, Wisconsin Institute for Discovery, University of Wisconsin - Madison, 330N Orchard Street, Madison, WI 53715, United States.
Abstract
The virus comet assay is a cell-based virulence assay used to evaluate an antiviral drug or antibody against a target virus. The comet assay differs from the plaque assay in allowing spontaneous flows in 6-well plates to spread virus. When implemented quantitatively the comet assay has been shown to have an order-of-magnitude greater sensitivity to antivirals than the plaque assay. In this study, a quantitative comet assay for influenza virus is demonstrated, and is shown to have a 13-fold increase in sensitivity to ribavirin. AX4 cells (MDCK cells with increased surface concentration of α2-6 sialic acid, the influenza virus receptor) have reduced the comet size variability relative to MDCK cells, making them a better host cell for use in this assay. Because of enhanced antiviral sensitivity in flow-based assays, less drug is required, which could lead to lower reagent costs, reduced cytotoxicity, and fewer false-negative drug screen results. The comet assay also serves as a readout of flow conditions in the well. Observations from comets formed at varying humidity levels indicate a role for evaporation in the mechanism of spontaneous fluid flow in wells.
Copyright ? 2011. Published by Elsevier B.V.
PMID:
22155578
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22155578