摘要
目的建立实时定量聚合酶链反应(RQ-PCR)方法并检测初诊急性早幼粒细胞白血病(APL)患者甲幼粒细胞白血病/维甲酸受体(PML/RARα)融合基因含量,评价检测结果的临床意义。方法采用"欧洲抗癌计划"推荐的引物和探针建立RQ-PCR方法,检测51例初诊APL患者的PML/RARα融合基因含量,并分析其临床意义。结果 APL初诊患者的年龄与外周血白细胞(WBC)数呈负相关(r=-0.376,P=0.014),相关性主要见于长型和变异型患者(r=-0.51,P=0.005)。51例APL初诊患者中,长型患者33例,变异型患者3例,长型和变异型36例。APL初诊患者PML/RARα转录本绝对含量为3.75×102～6.20×105拷贝/50ng(中位7.24×103拷贝/50ng);相对含量为0.51%～676.87%(中位9.31%)。15例短型患者PML/RARα转录本绝对含量为2.26×103～2.33×106拷贝/50ng(中位1.17×105拷贝/50ng);相对含量为12.04%～802.68%(中位95.26%)。短型患者外周血WBC数明显高于长型患者(P=0.041),其融合基因含量也明显高于长型患者(P<0.01),而在性别、年龄、外周血Hb、PLT,骨髓原粒及异常早幼粒细胞比例、血液学缓解时间和分子生物学缓解时间等方面比较差异无统计学意义。33例初诊患者血液学缓解时间、分子生物学缓解时间与患者的年龄、WBC数及融合基因含量无相关性。结论 RQ-PCR方法敏感、可靠,可以对APL初诊患者进行诊断和分型。
Objective To establish the real time quantitative PCR（RQ-PCR） assay and analyze its results in detection and quantification PML/RARα fusion transcripts in newly diagnosed patients with acute promyelocytic leukemia（APL）.Methods Three pairs of primer and TaqMan probe were designed according to ′Europe Against Cancer′（EAC） program for detecting the most frequent isoforms（Long-form,Short-form and Variant-form） of PML/RARα transcripts.To evaluate the utility of this assay,bone marrow samples from 51 newly diagnosed APL patients were detected and the results were analyzed.Results There was significant negative correlation between initial WBC count and the age of APL patients（r=-0.376,P=0.014）,mainly seen in the patients with Long-form and the Variant-form（r=-0.51,P=0.005）.In 51 newly diagnosed APL patients,there were 33 cases with Long-form PML/RARα transcripts,3 with Variant-form PML/RARα transcripts and 15 with Short-form PML/RARα transcripts.In 36 cases with Long-form or Variant-form transcripts,the median absolute and normalized amount of PML/RARα transcripts were 3.75×102-6.20×105 copies/50 ng（median 7.24×10^3 copies/50 ng） and 0.51%-676.87%（median 9.31%） respectively.In 15 cases with Short-form transcripts,the median absolute and normalized amount of PML/RARα transcripts were 2.26×103-2.33×106 copies/50 ng（median 1.17×10^5 copies/50 ng） and 12.04%-802.68%（median 95.26%） respectively.The WBC count was higher in the Short-form group than in the Long-form group（P=0.041）.The normalized amount of Short-form PML/RARα transcripts was significantly higher than that of Long-form（P0.01）.However,there was no statistical difference in the sex,age,Hb,PLT,proportion of the blast and promyelocyte in the bone marrow,the time of achieving hematologic complete remission and molecular complete remission between the Long-form patients and the Short-form patients.Among 33 cases with follow-up data,neither of the time of achieving hematologic complete remission nor molecular complete remission was correlated with the age,initial WBC and the expression level of the PML/RARα transcripts.Conclusion RQ-PCR for detecting the isoforms of PML/RARα fusion transcripts is a sensitive,reliable quantitative assay and can be used in the diagnosis of APL.
出处
《重庆医学》
CAS
CSCD
北大核心
2012年第1期1-3,7,共4页
Chongqing medicine
基金
江苏省医学重点人才基金资助项目(RC2007035)
江苏省自然基金资助项目(BK2009206)
镇江市社会发展基金资助项目(SH2008041)
关键词
聚合酶链反应
白血病
早幼粒细胞
急性
受体
维甲酸
polymerase chain reaction
leukemia
promyelocytic
acute
receptors
retinoicacid
