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J Immunol Methods. 2016 Jan 7. pii: S0022-1759(16)30006-0. doi: 10.1016/j.jim.2016.01.005. [Epub ahead of print]
Development of a multiplex quantitative PCR assay for the analysis of human cytokine gene expression in influenza A virus-infected cells.
Plotnikova MA1, Klotchenko SA2, Vasin AV3.
Author information
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.
Copyright ? 2015. Published by Elsevier B.V.
KEYWORDS:
Cytokines; Influenza virus; Multiplex quantitative real-time PCR
PMID: 26772136 [PubMed - as supplied by publisher]
Development of a multiplex quantitative PCR assay for the analysis of human cytokine gene expression in influenza A virus-infected cells.
Plotnikova MA1, Klotchenko SA2, Vasin AV3.
Author information
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.
Copyright ? 2015. Published by Elsevier B.V.
KEYWORDS:
Cytokines; Influenza virus; Multiplex quantitative real-time PCR
PMID: 26772136 [PubMed - as supplied by publisher]