Virus-inducible reporter genes as a tool for detecting and quantifying influenza A virus replication

Andrew Lutz, Julie Dyall, Paul D. Olivo, Andrew Pekosz

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)13-20
Number of pages8
JournalJournal of Virological Methods
Volume126
Issue number1-2
DOIs
StatePublished - Jun 2005
Externally publishedYes

Keywords

  • Cell culture
  • Diagnostics
  • Influenza
  • Reporter gene

ASJC Scopus subject areas

  • Virology

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