实时定量RT-PCR检测慢性粒细胞白血病bcr/abl融合基因的临床意义  

Application of real-time RT-PCR to detect bcr/abl fusion gene in chronic myeloid leukemia

作　　者：马晓霞[1] 王椿[1] 秦尤文[1] 颜式可[1] 高彦荣[1] 蔡琦[1] 

机构地区：[1]上海市第一人民医院血液科,上海200080

出　　处：《临床血液学杂志》2005年第3期164-168,共5页Journal of Clinical Hematology

摘　　要：目的:建立实时定量RTPCR方法检测慢性粒细胞白血病(CML)bcr/ablmRNA的方法;探讨CMLbcr/abl融合基因的表达水平与疗效的关系。方法:可同时检测b2a2和b3a2两种亚型,GAPDH的mRNA作为内参照。检测14例CML患者的22个外周血和骨髓样本中bcr/abl融合基因表达水平,并对2例异基因造血干细胞移植后复发的患者进行动态监测。结果:bcr/abl及GAPDH标准曲线的相关系数均为0.999;灵敏度为10-6;14例初诊患者的mRNA表达水平范围为2.81～145;2例异基因造血干细胞移植后复发患者的bcr/ablmRNA表达水平随临床治疗而变化。结论:实时定量PCR检测CML患者的bcr/abl融合基因,敏感可靠,重复性好;其融合基因表达水平的改变与临床疾病进展关系相一致,有助于反映白血病细胞负荷、评价疗效及判断疾病预后。Objective:To set up a real-time RT-PCR approach using TaqMan technology for detection and quantification of bcr/abl transcripts in CML patients and to explore the relationship between the expression level of the bcr/abl fusion transcript and the clinical status and efficacy of the therapy in CML.Method:Real-time PCR was carried out with single set of PCR primers and a probe allowing the detection of both b3a2 and b2a2 transcripts. The mRNA encoding for glyceraldehydes-3-phosphate dehydrogenase (GAPDH) was used as an endogenous reference. Twenty-two samples of bone marrow and peripheral blood from 14 CML patients were analyzed, and the bcr/abl mRNA levels of 2 HSCT patients after relapse were observed over time.Result:Both of the correlations of bcr/abl and GAPDH standard curves were 0.999; the sensitivity was 10 -6. The amounts of bcr/abl mRNA from 14 patients at diagnosis ranged from 2.81 to 145. The expression level of bcr/abl mRNA changed with the clinical outcome.Conclusion:This real-time RT-PCR is a reliable, sensitive and reproducible method of monitoring CML patients after therapy. It is useful in evaluating leukemic burden, assessing response to treatment and predicating the prognosis of the disease.

关 键 词：白血病 髓细胞性 实时定量聚合酶链反应 融合基因 

分 类 号：R733.72[医药卫生—肿瘤]