title: Fluorescence-based antiviral assay for the evaluation of compounds against vaccinia virus, varicella zoster virus and human cytomegalovirus.
abstract: Recombinant vaccinia virus (VACV), varicella zoster virus (VZV) and two human cytomegaloviruses (HCMV) expressing the green fluorescent protein (GFP) and the enhanced yellow fluorescent protein (EYFP) were used to develop a fluorescence-based assay for testing antiviral compounds. Infection of human embryonic lung fibroblasts (HEL) with the different recombinant viruses produced stable and detectable amount of GFP and EYFP signal as quantitated by automated fluorometry. The sensitivity of the recombinant viruses to a panel of antiviral drugs was measured and the fluorescence-based assay was compared to the cytopathic effect reduction assay (CPE-RA) in case of VACV and HCMV or to the plaque reduction assay (PRA) in case of VZV. The 50% inhibitory concentration (IC(50)) values for reference anti-pox and anti-herpesvirus compounds were comparable to those determined by CPE-RA or PRA assays. Furthermore the fluorimetric data could be confirmed by a flow cytometry assay. GFP- and EYFP-recombinant viruses proved to be a convenient tool for the evaluation of antiviral agents.
authors.affiliation: Rega Institute for Medical Research, K.U. Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium. fabiana.dalpozzo@libero.it
authors.forename: Fabiana
authors.initials: F
authors.lastname: Dal Pozzo
authors.forename: Graciela
authors.initials: G
authors.lastname: Andrei
authors.forename: Dirk
authors.initials: D
authors.lastname: Daelemans
authors.forename: Michael
authors.initials: M
authors.lastname: Winkler
authors.forename: Jacques
authors.initials: J
authors.lastname: Piette
authors.forename: Erik
authors.initials: E
authors.lastname: De Clercq
authors.forename: Robert
authors.initials: R
authors.lastname: Snoeck
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D003587:Cytomegalovirus / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D014645:Herpesvirus 3, Human / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D011995:Recombination, Genetic; D015203:Reproducibility of Results; D014616:Vaccinia virus / Q000187:drug effects* / Q000235:genetics / Q000502:physiology
chemical_list: D000998:Antiviral Agents; D049452:Green Fluorescent Proteins