title: A novel Ebola virus expressing luciferase allows for rapid and quantitative testing of antivirals.
abstract: Ebola virus (EBOV) causes a severe hemorrhagic fever with case fatality rates of up to 90%, for which no antiviral therapies are available. Antiviral screening is hampered by the fact that development of cytopathic effect, the easiest means to detect infection with wild-type EBOV, is relatively slow. To overcome this problem we generated a recombinant EBOV carrying a luciferase reporter. Using this virus we show that EBOV entry is rapid, with viral protein expression detectable within 2 h after infection. Further, luminescence-based assays were developed to allow highly sensitive titer determination within 48 h. As a proof-of-concept for its utility in antiviral screening we used this virus to assess neutralizing antibodies and siRNAs, with significantly faster screening times than currently available wild-type or recombinant viruses. The availability of this recombinant virus will allow for more rapid and quantitative evaluation of antivirals against EBOV, as well as the study of details of the EBOV life cycle.
authors.affiliation: Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT, USA. Electronic address: thomas.hoenen@nih.gov.
authors.forename: Thomas
authors.initials: T
authors.lastname: Hoenen
authors.forename: Allison
authors.initials: A
authors.lastname: Groseth
authors.forename: Julie
authors.initials: J
authors.lastname: Callison
authors.forename: Ayato
authors.initials: A
authors.lastname: Takada
authors.forename: Heinz
authors.initials: H
authors.lastname: Feldmann
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D001681:Biological Assay / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D029043:Ebolavirus / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter / Q000187:drug effects; D019142:Hemorrhagic Fever, Ebola / Q000188:drug therapy / Q000821:virology*; D006801:Humans; D008156:Luciferases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism
chemical_list: D000998:Antiviral Agents; D008156:Luciferases
keywords: Antiviral screening; CPE; EBOV; Ebola virus; Firefly; High-throughput screening; LBT assay; Luciferase; MOI; RLU; Reverse genetics; TCID(50); codon-optimized Firefly luciferase; cytopathic effect; eGFP; enhanced green fluorescent protein; luc2; luminescence-based direct titration assay; multiplicity of infection; relative light units; tissue culture infectious dose 50