摘要
目的:探讨实时定量RT-PCR方法检测急性早幼粒细胞白血病(APL)PML/RARα融合基因的临床意义。方法:采用实时定量RT-PCR方法,用Taqman探针检测32例初治APL患者的PML/RARα融合基因3种异构体表达水平,并对治疗过程中的20例患者融合基因表达水平进行动态观察。结果:①实时定量RT-PCR的敏感度为101拷贝数/μl,标准品日间差异及日内差异平均变异系数均<5%;②32例PML/RARα融合基因阳性的初治APL患者中,21例为PML/RARα融合基因长型异构体,11例为PML/RARα融合基因短型异构体;③32例初治患者PML/RARα融合基因表达量中位数和x-±s分别为1.44%,(1.29±1.46)%。比较异构体长型及短型标准拷贝数(NCN)中位数和x-±s分别为1.40%,(1.46±1.18)%和1.28%,(1.39±1.51)%,差异无统计学意义P<0.05;④20例治疗后APL患者PML/RARα融合基因表达水平随着临床治疗而改变。结论:①建立了检测APL微小残留病的高敏感性、高特异性的实时定量RT-PCR方法;②32例初治APL患者PML/RARα融合基因长型及短型异构体mRNANCN差异无统计学意义;③PML-RARα融合基因表达水平的变化与临床疾病发展相一致,有助于疗效评价、微小残留病的检测和预后判断。
Objective：To investigate the clinical value of quantification of PML/RARα fusion gene transcripts using real-time reverse transcription PCR approach. Method： Real-time RT-PCR was carried out to detect the type and the quantification of PML/RARα fusion gene transcripts in APL patients during the diagnose and therapy. Result： The sensitivity of real-time RT-PCR was 10^1 copies/ul. The coefficiency of interassay and intraassay was below 5%. Among 32 cases of APL patients with PML/RARa fusion gene positive, 21 were found with PML/ RARα L-type isoform, while 11 with S-type isoform. The median normalized copy number （NCN） of PML/RARα was 1.44% （1.29%±1.46%） in 32 APL patients at diagnosis. There was no significant difference of NCN between the two isoform groups. The expression level of PML/RARa fusion gene changed with the clinical outcome. Conclusion： The real-time quantitative RT-PCR is a sensitive approach for quantifying PML/RARa transcripts in APL patients. It is useful in assessing response to treatment and predicating the prognosis of the disease.
出处
《临床血液学杂志》
CAS
2008年第3期250-254,共5页
Journal of Clinical Hematology
基金
深圳市卫生局科技重点项目(No:200639)
