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目的对比肾移植患者和常规手术患者免疫状态,验证以β-actin mRNA为模板参照定量PCR技术的有效性。方法采用SYBR GreenⅠ荧光定量PCR对29例肾移植患者和10例常规手术患者术前、术后3和7天外周血淋巴细胞IL-2和IL-10 mRNA表达进行检测。结果两组术前IL-2和IL-10 mRNA水平无差别;术后肾移植组IL-2mRNA明显降低,而对照组无明显变化;两组术后第3天IL-10 mRNA表达水平均明显升高,肾移植组高于对照组(P<0.01),术后第7天对照组降至正常,肾移植组亦下降,但水平仍高于术前。结论SYBR GreenⅠ荧光定量PCR技术对于监测肾移植患者免疫状况和研究免疫耐受意义重大,以β-actin mRNA为模板参照的定量PCR技术是测定细胞因子基因表达的简单有效方法。
OBJECTIVE: To compare the immunological status of renal transplant recipients and patients undergoing routine surgery, and to verify the effectiveness of quantitative PCR using β-actin mRNA as a template. Methods The expression of IL-2 and IL-10 mRNA in peripheral blood lymphocytes of 29 renal transplant recipients and 10 routine surgical patients before and 3 and 7 days after operation were detected by SYBR GreenⅠ fluorescence quantitative PCR. Results There was no difference in IL-2 and IL-10 mRNA levels between the two groups before operation. IL-2 mRNA in renal transplantation group was significantly lower than that in control group after operation (P <0.01). The control group was reduced to normal on the 7th day after operation and the renal transplantation group was also decreased, but the level was still higher than that before operation. Conclusion SYBR Green Ⅰ fluorescence quantitative PCR is very important for monitoring immune status and immune tolerance in renal allograft recipients. Quantitative PCR with β-actin mRNA as a template is a simple and effective method for the determination of cytokine gene expression.