Objective To investigate the clinical significance of one step RQ-RT-PCR in detection of PML-RARα fusion gene on diagnose and minimal residual disease(MRD)in acute promyelocytic leukemia( APL). Methods One step RQ-RT-PCR and Taqman probe technique was used to detect two common types of PML-RARαfusion gene transcripts. Bone marrow samples from 42 APL and 30 APL-CR patients were analyzed. At the same time, Flow Cytometry( FCM) was used to monitor the MRD of bone marrow samples from some of the APL-CR patients. The consistency of two monitoring methods were compared. Results ①Among 42 APL patients, 34 cases were found with PML-RARa fusion gene L-type isoform, while 8 cases with S-type isoform. The median normalized copy number ( NCN) of 42 APL patients was 61% ( 13% ~ 284% ) , the median NCN of L-type isoform was 56% (28% ~ 278% ) , while that of S-type isoform was 70% ( 13% ~ 284% ) , there was no significant difference (P > 0.05) ; ②Bone marrow samples from 30 APL-CR patients were monitored consecutively on PML-RARαfusion gene by one step RQ-RT-PCR. The median NCN was 4.00% (0.00% ~8. 26% ); ③ Detecting MRD of APL-CR patients by one step RQ-RT-PCR and FCM at the same time, the two methods had high consistency(x2 =0.278, P = 0.598 ). The total coincidence rate was 83.3%. Conclusions One step RQ-RT-PCR is a simple and rapid method that has high specificity and sensitivity, which has an important clinical significance on diagnose and monitor MRD in APL.%目的 探讨一步法实时定量RT-PCR (RQ-RT-PCR)技术在急性早幼粒细胞白血病(APL)患者诊断及微小残留病(MRD)检测中的临床价值.方法 应用一步法RQ-RT-PCR、Taqman探针技术检测42例APL初诊患者和30例完全缓解(CR)患者骨髓PML-RARo融合基因的两种常见异构体转录本mRNA的表达.同时采用多参数流式细胞术(FCM)对部分患者骨髓MRD进行检测,并比较两种检测方法的一致性.结果 ①42例APL初诊患者的PML-RARα融合基因转录本中位标准拷贝数(NCN)为61％(13％ ～284％),其中L型34例,S型8例.L型和S型异构体中位NCN分别为56％(28％ ～ 278％)和70％(13％ ～284％),两者比较,P＞0.05；②30例APL患者经标准诱导治疗CR后PML-RARα融合基因转录本中位NCN为4.00％(0.00％ ～ 8.26％)；③一步法RQ-RT-PCR和FCM同时检测结果有较好的一致性(x2=0.278,P=0.598),总符合率83.3％.结论 一步法RQ-RT-PCR具有操作简便、特异性好、灵敏度高、结果可靠、直观等特点,有利于对APL的诊断和MRD检测.
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