实时荧光定量PCR方法检测非霍奇金淋巴瘤患者B淋巴细胞刺激因子表达水平

摘要

目的:建立实时荧光定量PCR方法(RFQ-PCR)检测非霍奇金淋巴瘤(NHL)患者B淋巴细胞刺激因子(BAFF)表达水平.方法:47例NHL患者及20例健康对照,采用实时荧光定量PCR方法检测其外周血单个核细胞中的B淋巴细胞刺激因子表达水平.结果:RFQ-PCR检测 BAFF含量的示灵敏度为10 pg/ml,低浓度样本批内和批间变异系数(CV)分别为8.56%和 11.32%,高浓度样本批内和批间CV分别为0.76%和4.58%.NHL患者和健康对照者 BAFF mRNA表达量分别为(0.48±0.023,0.25±0.023),两者具有显著性差异(P=0.0001).结论:RFQ-PCR检测BAFF mRNA含量的方法,具有较好的检测灵敏度和重复性.NHL患者BAFF mRNA 高表达,提示BAFF可能在NHL发生发展中发挥重要作用.%Objective:To establish the examination method of BAFF mRNA expression in non-Hodgkin lymphoma peripheral blood mononuclear cells by real-time fluorescence quantitative PCR. Methods: The BAFF mRNA expression of 47 non-Hodgkin lymphoma (NHL) patients and 20 healthy peripheral blood mononuclear cells were examined by real-time fluorescence quantitative PCR ( RFQ-PCR) . Results:The sensitivity of RFQ-PCR was 10 pg/ml. The variation coefficients of intra-assay and inter-assay for low concentration sample were 8. 56% and 11. 32% , for high concentration sample were 0.76% and 4. 58% respectively. The expression of BAFF mRNA in NHL patients were significantly higher compared to those in control groups( 0.48 ±0. 023,0. 25 ± 0. 023) . Conclusion; This assay has high sensitivity and reproducibility. It is suitable for measurement of the expression level of BAFF. The BAFF mRNA expression in NHL patients peripheral blood were higher compared to those in control groups, which may play important roles in the initiation and progression of NHL.