实时定量RT-PCR检测PML-RARα融合基因诊断APL的临床价值  

Real-time quantitative RT-PCR in detection of PML-RARαfusion gene in APL

作　　者：陆伟[1] 秦燕[1] 丁润生[1] 尹红[1] 

机构地区：[1]南通大学附属医院,江苏南通226001

出　　处：《山东医药》2011年第33期13-15,共3页Shandong Medical Journal

摘　　要：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-RARα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(χ2=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。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-RARα 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%）;③Detecting MRD of APL-CR patients by one step RQ-RT-PCR and FCM at the same time,the two methods had high consistency（χ2=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.

关 键 词：白血病 早幼粒细胞 急性 PML-RARΑ融合基因 实时定量RT-PCR 微小残留病 

分 类 号：R557[医药卫生—血液循环系统疾病]