Development of a multiplex quantitative PCR assay for the analysis of human cytokine gene expression in influenza A virus-infected cells

Journal of Immunological Methods
Marina A PlotnikovaAndrey V Vasin

Abstract

Cytokines are global mediators of cellular communications that are involved in broad array of biological processes, including the immunological and inflammatory mechanisms of virus-host interactions. Measuring the gene expression of simultaneously expressed cytokines is necessary for understanding the pathogenesis of many viral infections, including influenza. We developed a multiplex quantitative real-time PCR (qPCR) method for the detection of the following human cytokines: IL-1B, IL-2, IL-4, IL-6, IL-10, IL-12B, IL-18, IFN-γ and TNF. The assay consisted of three sets of multiple qPCRs; in each qPCR, three target cytokines and reference GAPDH genes were amplified. The assay provided a precise and sensitive quantification of cytokine gene expression with a 20fmol limit of detection and a 1.5% coefficient of variation. This method was successfully applied to cytokine profiling in epithelial A549 cells that were infected with A/California/07/09 (H1N1pdm2009) virus.

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Citations

Jun 12, 2016·Toxicology and Applied Pharmacology·Kuo-Liang WeiJyan-Gwo Joseph Su
Apr 11, 2015·Comparative Immunology, Microbiology and Infectious Diseases·Cláudia S MarquesGabriela M Santos-Gomes
May 7, 2014·Trends in Microbiology·Marie GerberRoland Marquet
Jun 12, 2021·Advanced Science·Chao LiuGuozhen Liu

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