实时定量聚合酶链反应动态监测白血病治疗后微小残留病研究

摘要

目的 建立急性早幼粒细胞白血病(APL)实时定量聚合酶链反应(RQ-PCR)检测方法,并检测APL治疗后微小残留病(MRD),评价MRD检测的意义.方法 应用RQ-PCR对96例APL患者的骨髓标本进行分析,检测PML/RARa融合基因的转录水平.结果 (1)RQ-PCR与传统巢式PCR(RT-PCR)检测结果的差异有统计学意义(P＜0.05),提示应用RQ-PCR检测PML/RARa融合基因转录水平的敏感性高于RT-PCR.(2)RQ-PCR可对PML/RARa融合基因进行定量检测,监测患者的PML-RARa融合基因拷贝数水平变化,结果显示初诊时较高,有效治疗后迅速下降,复发时又出现上升,提示若患者在完全缓解(CR)后PML/RARa融合基因拷贝数呈现上升趋势,其复发机率大.结论 RQ-PCR检测作为APL治疗后微小残留病检测的方法,准确可靠,可以判断治疗效果,预测疾病复发,早期给予干预治疗,有重要的临床应用价值.%Objective To set up a real-time quantitative PCR approach for detection and quantification for PML/ RARa transcripts in APL patients, and detect minimal residual disease (MRD) in APL and evaluate the significance of MRD detection. Method Application of real-time quantitative PCR in bone marrow specimens for analysis of 96 patients with APL, detection of PML/RARa fusion gene transcription. Results Application of rank-sum test on the real-time PCR and nested PCR sensitivity analysis, the results showed statistically significant(P<0. 05). Prompt application of real-time quantitative PCR detection of PML/RARa fusion gene transcript levels, the sensitivity was higher than the traditional nested PCR. And real-time quantitative PCR could provide a specific value. easy to describe the level changes of patients with PML/RARa fusion gene copy number. The newly diagnosed was high and rapidly decreased after effective treatment, reappeared after recarrence, suggesting that if the patients in complete remission (CR) with PML / RARa fusion gene copy number on the rise, the probability of relapse increased. Conclusion Real-time quantitative PCR is reliable and has important clinical value, it can be used to detect the MRD, monitor the treatment outcome, predict the disease recurrence and give early intervention.