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Influenza virus assays based on virus-inducible reporter cell lines

机译:基于病毒诱导的报告细胞系的流感病毒检测

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Background 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. Findings 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. Conclusions 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.
机译:背景技术病毒诱导的报告基因已被用作许多病毒的病毒检测和定量分析的基础。最近描述了一种用于报告基因的甲型流感病毒诱导的策略。在本报告中,我们描述了这种策略对乙型流感病毒的扩展,具有甲型和乙型流感病毒可诱导的报告基因的稳定细胞系的产生,以及这些细胞在各种临床相关病毒测定中的用途。本文所述的每种细胞系组成性地表达RNA转录物,该转录物包含报告基因编码区,该报告基因编码区的侧翼是病毒的5'非翻译区和3'非翻译区(UTR),因此模拟流感病毒基因组区段。流感病毒感染细胞后,病毒诱导的报告基因片段(VIRGS)被病毒聚合酶复合物复制并转录,从而导致报告基因表达。 Findings Reporter基因的诱导是在感染了许多实验室株和包括几株H5N1株的流感病毒临床分离株后发生的。诱导是剂量依赖性的,并且对甲型或乙型流感病毒具有高度特异性。结论这些细胞系为涉及病毒定量的简单,快速和客观的测定提供了基础,例如病毒滴度的测定,抗病毒药性的评估以及抗体中和滴度的测定。这些细胞系对于流感病毒研究人员和疫苗制造商可能非常有用。

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