Peer-reviewed veterinary case report
Establishment of real-time TaqMan-fluorescence quantitative RT-PCR assay for detection and quantitation of ten kinds of porcine inflammation markers mRNA.
- Journal:
- Journal of virological methods
- Year:
- 2013
- Authors:
- Lin, Wencheng et al.
- Affiliation:
- Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences · China
Plain-English summary
Researchers developed a new test to measure certain inflammation markers in pigs. This test is very precise and can detect tiny amounts of these markers, which are important for understanding how pigs respond to inflammation. They found that when they treated pig testicle cells with a specific substance, some inflammation markers showed increased activity, while others did not change much. This new testing method could help scientists better understand inflammation in pigs and potentially improve their health management.
Abstract
Real-time PCR assays based on TaqMan probes for detection of porcine inflammation markers including interferon-α (IFN-α), interferon-β (IFN-β), 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 × 10(1)copies/μl, 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 β-actin in swine testicle (ST) cells transfected with polyinosinic: polycytidylic acid (poly (I:C)). The results showed that the transcription levels (mRNA) of IFN-α, IFN-β, 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.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/23792113/