tetano
Editor, Senior Moderator
J Immunol Methods. 2016 Jan 7. pii: S0022-1759(16)30006-0. doi: 10.1016/j.jim.2016.01.005. [Epub ahead of print]
[h=1]Development of a multiplex quantitative PCR assay for the analysis of human cytokine gene expression in influenza A virus-infected cells.[/h] Plotnikova MA[SUP]1[/SUP], Klotchenko SA[SUP]2[/SUP], Vasin AV[SUP]3[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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 (H1N1[SUB]pdm2009[/SUB]) virus.
Copyright ? 2015. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] Cytokines; Influenza virus; Multiplex quantitative real-time PCR
PMID: 26772136 [PubMed - as supplied by publisher]
[h=1]Development of a multiplex quantitative PCR assay for the analysis of human cytokine gene expression in influenza A virus-infected cells.[/h] Plotnikova MA[SUP]1[/SUP], Klotchenko SA[SUP]2[/SUP], Vasin AV[SUP]3[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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 (H1N1[SUB]pdm2009[/SUB]) virus.
Copyright ? 2015. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] Cytokines; Influenza virus; Multiplex quantitative real-time PCR
PMID: 26772136 [PubMed - as supplied by publisher]