期刊
JOURNAL OF VIROLOGICAL METHODS
卷 126, 期 1-2, 页码 13-20出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2005.01.016
关键词
influenza; diagnostics; reporter gene; cell culture
资金
- NIAID NIH HHS [R01 AI053629-01A1, R01 AI053629, R01 AI053629-02] Funding Source: Medline
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 transcript, 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. (c) 2005 Elsevier B.V. All rights reserved.
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