Diagnostic Hybrids Inc., Athens, OH, USA.
Influenza Other Respir Viruses. 2009 Sep;3(5):241-51. doi: 10.1111/j.1750-2659.2009.00095.x.
Virus-inducible reporter genes have been used as the basis of virus detection and quantitation assays for a number of viruses. A strategy for influenza A virus-induction of a reporter gene was recently described. In this report, we describe the extension of this strategy to influenza B virus, the generation of stable cell lines with influenza A and B virus-inducible reporter genes, and the use of these cells in various clinically relevant viral assays. Each of the cell lines described herein constitutively express an RNA transcript that contains a reporter gene coding region flanked by viral 5￠- and 3￠-untranslated regions (UTR) and therefore mimics an influenza virus genomic segment. Upon infection of the cells with influenza virus the virus-inducible reporter gene segment (VIRGS) is replicated and transcribed by the viral polymerase complex resulting in reporter gene expression.
Reporter gene induction occurs after infection with a number of laboratory strains and clinical isolates of influenza virus including several H5N1 strains. The induction is dose-dependent and highly specific for influenza A or influenza B viruses.
These cell lines provide the basis of simple, rapid, and objective assays that involve virus quantitation such as determination of viral titer, assessment of antiviral susceptibility, and determination of antibody neutralization titer. These cell lines could be very useful for influenza virus researchers and vaccine manufacturers.
病毒诱导型报告基因已被用作多种病毒的病毒检测和定量分析的基础。最近描述了一种用于甲型流感病毒诱导报告基因的策略。在本报告中，我们将该策略扩展到乙型流感病毒，生成具有甲型和乙型流感病毒诱导型报告基因的稳定细胞系，并将这些细胞用于各种临床相关的病毒检测。本文所述的每个细胞系均稳定表达一种 RNA 转录本，该转录本包含侧翼为病毒 5' 和 3' 非翻译区（UTR）的报告基因编码区，因此模拟了流感病毒基因组片段。当细胞被流感病毒感染时，病毒诱导型报告基因片段（VIRGS）被病毒聚合酶复合物复制和转录，导致报告基因表达。
报告基因诱导发生在感染多种实验室株和临床分离株的流感病毒后，包括几种 H5N1 株。诱导是剂量依赖性的，对甲型或乙型流感病毒具有高度特异性。
这些细胞系为涉及病毒定量的简单、快速和客观的检测提供了基础，例如测定病毒滴度、评估抗病毒敏感性和测定抗体中和滴度。这些细胞系对于流感病毒研究人员和疫苗制造商非常有用。
