title: A colorimetric bioassay for high-throughput and cost-effectively assessing anti-foot-and-mouth disease virus activity.
abstract: Foot-and-mouth disease virus (FMDV) is one of the most contagious animal viruses. This virus is very sensitive to inhibition by type I interferons. Currently, a bioassay based on plaque reduction is used to measure anti-FMDV activity of porcine IFNs. The plaque reduction assay is tedious and difficult to utilize for high-throughput analysis. Using available FMDV susceptible bovine and porcine cells, we developed and tested a colorimetric assay based on cytopathic effect reduction for its ability to quantify FMDV-specific antiviral activity of bovine and porcine type I interferons. Our results show that this new method has significant advantages over other assays in terms of labor intensity, cost, high-throughput capability and/or anti-FMDV specific activity because of simpler procedures and direct measurement of antiviral activity. Several assay conditions were tested to optimize the procedures. The test results show that the assay can be standardized with fixed conditions and a standard or a reference for measuring antiviral activity as units. This is an excellent assay in terms of sensitivity and accuracy based on a statistical evaluation. The results obtained with this assay were highly correlated with a conventional virus titration method.
authors: Foreign Animal Disease Research Unit, Agricultural Research Unit, United States Department of Agriculture, Plum Island Animal Disease Research Center, Orient Point, NY 11957, USA.
authors: Palaniappan
authors: P
authors: Ramanathan
authors: Foreign Animal Disease Research Unit, Agricultural Research Unit, United States Department of Agriculture, Plum Island Animal Disease Research Center, Orient Point, NY 11957, USA. Electronic address: james.zhu@ars.usda.gov.
authors: James J
authors: JJ
authors: Zhu
authors: Foreign Animal Disease Research Unit, Agricultural Research Unit, United States Department of Agriculture, Plum Island Animal Disease Research Center, Orient Point, NY 11957, USA.
authors: Elizabeth A
authors: EA
authors: Bishop
authors: Foreign Animal Disease Research Unit, Agricultural Research Unit, United States Department of Agriculture, Plum Island Animal Disease Research Center, Orient Point, NY 11957, USA.
authors: Michael C
authors: MC
authors: Puckette
authors: Foreign Animal Disease Research Unit, Agricultural Research Unit, United States Department of Agriculture, Plum Island Animal Disease Research Center, Orient Point, NY 11957, USA.
authors: Ethan
authors: E
authors: Hartwig
authors: Foreign Animal Disease Research Unit, Agricultural Research Unit, United States Department of Agriculture, Plum Island Animal Disease Research Center, Orient Point, NY 11957, USA.
authors: Marvin J
authors: MJ
authors: Grubman
authors: Foreign Animal Disease Research Unit, Agricultural Research Unit, United States Department of Agriculture, Plum Island Animal Disease Research Center, Orient Point, NY 11957, USA. Electronic address: luis.rodriguez@ars.usda.gov.
authors: Luis L
authors: LL
authors: Rodriguez
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000191:economics / Q000379:methods / Q000662:veterinary*; D002417:Cattle; D002460:Cell Line; D003124:Colorimetry / Q000191:economics / Q000379:methods / Q000662:veterinary*; D003362:Cost-Benefit Analysis; D003588:Cytopathogenic Effect, Viral / Q000276:immunology; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology* / Q000472:pathogenicity*; D057166:High-Throughput Screening Assays / Q000191:economics / Q000379:methods / Q000662:veterinary; D007113:Immunity, Innate; D007370:Interferon Type I / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D034421:Sus scrofa
chemical_list: D007370:Interferon Type I; D011994:Recombinant Proteins
keywords: Antiviral activity; Cytopathic effect reduction assay; Foot-and-mouth disease virus; Type I interferons