实时定量RT—PCR方法检测初诊白血病患者常见分子标志物  

Determination of the common molecular markers in newly diagnosed leukemias by using real-time quantitative RT-PCR

作　　者：姚利[1] 陈子兴[1] 岑建农[1] 邱桥成[1] 何军[1] 鲍晓晶[1] 袁晓妮[1] 

机构地区：[1]苏州大学附属第一医院、江苏省血液研究所,215006

出　　处：《中华血液学杂志》2008年第3期192-195,共4页Chinese Journal of Hematology

基　　金：国家863计划资助项目（2006AA02A405）

摘　　要：目的建立实时定量RT-PCR（RQ—RT—PCR）方法，检测初诊白血病患者骨髓细胞中的常见特征性分子生物学标志物的表达水平，评价其在疾病诊断和微量残留病（MRD）监测中的意义。方法设计TaqMan探针和引物，建立RQ-RT-PCR法对常见融合基因转录本（bcr-abl、AML-ETO、PML-RARer）和abl阳性模板进行扩增，检测202例初诊白血病患者的融合基因转录本含量。结果初诊白血病患者中的融合基因表达水平以绝对量表示，bcr—abl转录本b3a2或b2a2型拷贝数为47614．63，bcr-abl转录本ela2型拷贝数为98847．53，AML1-ETO转录本拷贝数为300029．51，PML-RARα转录本拷贝数为25506．28。以abl校正拷贝数（NCN）表示相对量，bcr-abl转录本b3a2或b2a2型为1．05、bcr-abl转录本ela2型为0．91、AML1-ETO转录本为5．33、PML-RARα转录本为0．55。结论以abl校正拷贝数计算融合基因表达水平，结果更准确、直观，优于绝对定量。同时，不同类型融合基因转录本在初诊白血病患者中表达水平不同。Objective To establish a real-time quantitative reverse transcriptase polymerase chain reaction （RQ-RT-PCR） for quantitative detection of the common molecular markers that have affirmative clinical significance in the acute and chronic leukemia patients, and evaluate its significance in diagnosing leukemias and monitoring minimal residual disease（MRD）. Methods Primers and TaqMan probes were designed for detecting various fusion transcripts and normal abl gene was used as the internal control. The expression level of fusion transcripts in 202 newly diagnosed leukemias were determined. Results In absolute quantity, expression level of the fusion transcripts in various leukemias was b3a2（b2a2） 47 614.63, ela2 98 847.53, AML1-ETO 300 029.51, PML-RARα 25 506.28, respectively, while in relative quantity to abl, the levels were 1.05, 0.91, 5.33 and 0.55, respectively. Conclusion The relative quantification of gene expression level by using RQ-RT-PCR to abl control gene is more accurate and direct viewing. Different levels of transcription of corresponding fusion genes are found in various subtypes of leukemias at diagnosis, among which the level of AML1-ETO was higher and PML-RARα lower.

关 键 词：逆转录聚合酶链反应 实时定量 白血病 融合基因转录本 

分 类 号：R686[医药卫生—骨科学]