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

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• 作者：Wencheng Lin ; Zhongtian Wang ; Zheng Qiu ; Qinfang Liu ; Shangjin Cui
• 关键词：Cytokine ; Porcine ; Real-time PCR ; Type I interferon ; Inflammation markers
• 刊名：Journal of Virological Methods
• 出版年：2013
• 出版时间：October, 2013
• 年：2013
• 卷：193
• 期：1
• 页码：169-172
• 全文大小：357 K

文摘

Real-time PCR assays based on TaqMan probes for detection of porcine inflammation markers including interferon-¦Á (IFN-¦Á), interferon-¦Â (IFN-¦Â), 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 ¡Á 10¹ copies/¦Ìl, 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 ¦Â-actin in swine testicle (ST) cells transfected with polyinosinic: polycytidylic acid (poly (I:C)). The results showed that the transcription levels (mRNA) of IFN-¦Á, IFN-¦Â, RIG-1 and IRF-7 were up-regulated in ST cells transfected with poly (I:C), 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.