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目的:建立荧光定量PCR检测三氯乙烯药疹样皮炎(DMLT)患者外周血淋巴细胞CXCR2和CXCR3mRNA表达的方法。方法:利用SYBRGreen荧光定量PCR分别检测24例DMLT患者、26例正常人外周血淋巴细胞CXCR2和CXCR3基因mRNA表达情况,以β2微球蛋白基因作为内参,根据相对定量公式(2-△△CT)计算DMLT患者与正常人CX-CR2和CXCR3基因表达差异倍数。结果:24例患者外周血淋巴细胞荧光定量PCR均检出CXCR2和CXCR3mRNA表达,其中CXCR2表达情况有两种:表达上升14例(58.3%)和表达下降10例(41.7%),与正常人CXCR2mRNA相比表达倍数分别为16.76±7.01、0.54±0.30;CXCR3表达显著升高(△CT=6.3±2.8,11.4±1.9;P<0.01),与正常人CX-CR3mRNA相比表达倍数为33.37±31.61。结论:成功建立了DMLT患者外周血淋巴细胞CXCR2和CXCR3mRNA表达的荧光定量PCR方法。
Objective: To establish a method for the detection of CXCR2 and CXCR3 mRNA expression in peripheral blood lymphocytes of patients with trichlorethylene eruptive dermatitis (DMLT) by fluorescence quantitative PCR. Methods: The mRNA expression of CXCR2 and CXCR3 in 24 cases of DMLT and 26 cases of normal human peripheral blood lymphocytes were detected by SYBR Green quantitative PCR. The β2 microglobulin gene was used as an internal control. According to the relative quantification formula (2- △△ CT) Calculate the difference between CXCR2 and CXCR3 gene expression in DMLT patients and normal subjects. Results: The expression of CXCR2 and CXCR3 mRNA in 24 peripheral blood lymphocytes were detected by fluorescence quantitative PCR. The expression of CXCR2 was detected in 14 patients (58.3%) and 10 (41.7%) in CXCR2, (△ CT = 6.3 ± 2.8, 11.4 ± 1.9; P <0.01), and the expression of CXCR3 was significantly higher than that of normal human CX-CR3 mRNA (33.37 ± 31.61 vs 16.76 ± 7.01,0.54 ± 0.30, respectively) . Conclusion: The fluorescence quantitative PCR method of CXCR2 and CXCR3 mRNA expression in peripheral blood lymphocytes of DMLT patients has been successfully established.