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| Establishment and application of the real-time fluorescence quantitative PCR assay for detection of human IL-21 mRNA expression |
| ZHU Chen-u, CHEN Yan-ong, WEI Yan-ai, QIN Wei, MA Jie, XU Hua-i, WANG Sheng-jun |
| (Institute of Laboratory Medicine, Jiangsu University, Zhenjiang Jiangsu 212013,China) |
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Abstract Objective: To establish realtime fluorescence quantitative PCR（qRT-PCR）method for detection of human interleukin-21（IL-21） mRNA expression. Methods：The peripheral blood mononuclear cells(PBMC) were separated and CD4+ cells were isolated using microbeads and stimulated by PHA. Total RNAs were extracted from CD4+ T cells and then transcribed reversely into cDNAs. The relative expression levels of human IL-21 mRNA in CD4+ T cells from patients with Graves′disease(GD） and healthy controls were detected by this method. The sensitivity and specificity of the method were evaluated. Results：The melting curve showed a single peak and amplified products were electrophoresed and a single product was detected. The IL-21 mRNA relative expression was significantly increased in GD patients compared to healthy controls(P＜0.001）. Conclusion：The method to detect IL-21 by qRT-PCR was good in specificity. It can be used as a standard method of qRT-PCR for detection of human IL-21 expression.
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Received: 01 June 2012
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2013, Vol. 23 
