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目的建立实时荧光定量PCR(RQ-PCR)检测融合基因TEL-AML1的方法,初步探讨其在儿童白血病临床诊治中的应用。方法采用Taqman探针技术建立RQ-PCR检测融合基因TEL-AML1的方法,利用该方法对24例TEL-AML1检测阳性的ALL患儿初发及随访骨髓标本进行微小残留病(MRD)定量分析,并与常规骨髓细胞形态学检查、流式细胞术(FCM)检测结果进行比较,探讨MRD定量信息与患儿治疗反应、临床预后等方面的相关性。结果诱导期末完全缓解(CR)患儿11例,经RQ-PCR检测5例(45%)MRD阳性;15例在维持治疗期间经骨髓细胞形态学检查均达CR,经RQ-PCR检测3例MRD阳性,经强化治疗和维持治疗后MRD水平均逐渐下降并最后转阴。复发1例患儿在其复发前1个月,经RQ-PCR检测MRD水平上升约103拷贝,予复发方案诱导化疗后再次达CR。结论RQ-PCR检测融合基因TEL-AML1的方法较常规骨髓细胞形态学方法、FCM具有更高的敏感度,可以定量检测MRD,为早期预测复发,指导临床治疗提供依据。
Objective To establish a real-time fluorescence quantitative PCR (RQ-PCR) method to detect the fusion gene TEL-AML1 and to explore its clinical application in the diagnosis and treatment of childhood leukemia. Methods TQ-PCR was used to detect the fusion gene TEL-AML1 by Taqman probe technique. The minimal invasive disease (MRD) quantitative analysis was performed in 24 newly diagnosed TEL patients with ALL-positive TEL-AML1 and the follow-up bone marrow samples by this method. And compared with conventional bone marrow cell morphology and flow cytometry (FCM) test results to explore the relationship between MRD quantitative information and children’s treatment response and clinical prognosis. Results Eleven patients with complete remission (CR) induced by induction were positive for MRD in 5 cases (45%) by RQ-PCR and 15 cases were all treated by morphological examination of bone marrow cells during maintenance therapy and 3 cases were detected by RQ-PCR MRD positive, after intensive treatment and maintenance therapy MRD levels were gradually decreased and finally turned negative. Recurrence 1 patient 1 month before the recurrence, RQ-PCR detection of MRD levels increased by about 103 copies, to relapse regimen after induction of chemotherapy CR again. Conclusion The method of RQ-PCR in detecting fusion gene TEL-AML1 is more sensitive than conventional bone marrow cell morphology method and FCM, and can quantitatively detect MRD. It provides the basis for early prediction of recurrence and clinical treatment.