Real-time quantitative RT-PCR detection of human peripheral blood mononuclear cells in the expression of FOXP3 mRNA.docVIP

 PAGE \* MERGEFORMAT 15 Real-time quantitative RT-PCR detection of human peripheral blood mononuclear cells in the expression of FOXP3 mRNA Of: Jiang Weiping, Dingmao Wen, ZHU Ya Fei, Li Bing, Zhu late Lin, Li Xinhua, Lin Feifei [Abstract] Objective: To establish a real-time quantitative reverse transcription - polymerase chain reaction (real-time fluorescence quantitative RT-PCR) detection of human peripheral blood mononuclear cells FOXP3 mRNA in the method, and study with CD4 + CD25 + Treg cell activity related . Methods: Peripheral blood mononuclear cells extracted from human total RNA, the mRNA reverse transcription into cDNA, the -actin as an internal reference, real-time quantitative RT-PCR detected 29 cases of asthmatic children and 24 cases were the same age the relative FOXP3 mRNA expression, the use of melting curves and agarose gel electrophoresis PCR product specificity, while use of flow cytometry CD4 + CD25 + Treg cell levels, correlation analysis between the two. Results: The asthma group CD4 + CD25 + Treg cells was significantly lower than the same age group, P lt;0.01, FOXP3 mRNA expression was also significantly lower, P lt;0.05, FOXP3 and -actin were melting curve analysis showed that only a single peak, Tm values #8203;#8203;were 82.4 and 87.8 , agar Sugar electrophoresis showed only a single PCR product are. Conclusion: SYBR Green real time quantitative RT-PCR expression levels of FOXP3 mRNA detected by simple and reliable results. preliminary results confirm that peripheral blood CD4 + CD25 + Treg cell number and FOXP3 mRNA expression of the same trend, there is a certain correlation. [Keywords:] real time quantitative RT-PCR, asthma, FOXP3, CD4 + CD25 + regulatory T cells Abstract: Objective: To establish a real-time fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCRto detect FOXP3 mRNA expression in human peripheral blood mononuclear cells (PBMCs and to investigate the correlations between acti