title: Virus-inducible reporter genes as a tool for detecting and quantifying influenza A virus replication.
abstract: The use of influenza A virus-inducible reporter gene segments in detecting influenza A virus replication was investigated. The RNA polymerase I promoter/terminator cassette was used to express RNA transcripts encoding green fluorescence protein or firefly luciferase flanked by the untranslated regions of the influenza A/WSN/33 nucleoprotein (NP) segment. Reporter gene activity was detected after reconstitution of the influenza A virus polymerase complex from cDNA or after virus infection, and was influenza A virus-specific. Reporter gene activity could be detected as early as 6 h post-infection and was virus dose-dependent. Inhibitory effects of antibodies or amantadine could be detected and quantified rapidly, providing a means of not only identifying influenza A virus-specific replication, but also of determining the antigenic subtype as well as antiviral drug susceptibility. Induction of virus-specific reporter genes provides a rapid, sensitive method for detecting virus replication, quantifying virus titers and assessing antiviral sensitivity as well as antigenic subtype.
authors.affiliation: Department of Molecular Microbiology and Pathology, Washington University School of Medicine, Campus Box 8230, 660 S. Euclid Avenue, St. Louis, MO 63110, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Lutz
authors.forename: Julie
authors.initials: J
authors.lastname: Dyall
authors.forename: Paul D
authors.initials: PD
authors.lastname: Olivo
authors.forename: Andrew
authors.initials: A
authors.lastname: Pekosz
mesh_terms: D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D004285:Dogs; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D009980:Influenza A virus / Q000235:genetics / Q000502:physiology*; D049409:Luciferases, Firefly / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D011401:Promoter Regions, Genetic; D012318:RNA Polymerase I / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D012703:Serotyping; D013194:Staining and Labeling / Q000379:methods; D020506:Untranslated Regions; D014779:Virus Replication
chemical_list: D012367:RNA, Viral; D020506:Untranslated Regions; D049452:Green Fluorescent Proteins; D049409:Luciferases, Firefly; D012318:RNA Polymerase I