Establishment of real-time TaqMan-fluorescence quantitative RT-PCR assay for detection and quantitation of ten kinds of porcine inflammation markers mRNA

文献类型: 外文期刊

第一作者: Lin, Wencheng

作者: Lin, Wencheng;Qiu, Zheng;Liu, Qinfang;Cui, Shangjin;Wang, Zhongtian

作者机构:

关键词: Cytokine;Porcine;Real-time PCR;Type I interferon;Inflammation markers

期刊名称:JOURNAL OF VIROLOGICAL METHODS ( 影响因子:2.014; 五年影响因子:2.001 )

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收录情况: SCI

摘要: Real-time PCR assays based on TaqMan probes for detection of porcine inflammation markers including interferon-alpha (IFN-alpha), interferon-beta (IFN-beta), retinoic acid-inducible gene 1 (RIG-1), toll-like receptor 9 (TLR-9), interferon regulatory factors (IRF-3, IRF-7), Janus kinase (JAK-1, JAK-2), signal transducers and activators of transcription (STAT-1, STAT-2) were established in this study. The results indicated that the established assays were highly specific and sensitive with a detection limit of 1.0 x 10(1) copies/mu l, and coefficient of variations was less than three percent for both inter- and intra-assay. The established assays were used to detect mRNA levels of these inflammation markers and beta-actin in swine testicle (ST) cells transfected with polyinosinic: polycytidylic acid (poly (I:C)). The results showed that the transcription levels (mRNA) of IFN-alpha, IFN-beta, RIG-1 and IRF-7 were up-regulated in ST cells transfected with poly (LC), and the transcription levels (mRNA) of TLR-9, IRF-3, JAK-1, JAK-2, STAT-1 and STAT-2 showed minimal change. The real-time PCR assays established in this study with high specificity, sensitivity and reproducibility could be used to quantify mRNA levels of porcine inflammation markers

分类号: R37

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