摘要
目的检测初治急性早幼粒细胞白血病(APL)患者PML/RARα融合基因异构体类型及表达水平,建立检测微小残留病的标准拷贝数基线。方法应用实时定量RT-PCR方法,采用Taqman探针技术测定32例APL初治患者骨髓标本的PML/RARα融合基因mRNA3种异构体表达水平,并比较异构体之间临床特征有无差异。结果32例PML/RARα融合基因阳性的APL初治患者中21例为PML/RARα融合基因长型异构体,11例为融合基因短型异构体,初治患者融合基因表达量中位数为1.44%(1.29%±1.46%)。长型及短型异构体标准拷贝数(normalized copy number,NCN)中位数分别为1.40%(1.46%±1.18%)和1.28%(1.39%±1.51%),无明显统计学差异,P<0.05,融合基因长型异构体患者在发病时外周血白细胞总数为6.08±13.21(×109/l)明显低短型患者15.11±20.26(×109/l),P<0.01。结论建立了检测APL微小残留病的高敏感性、高特异性的实时定量RT-PCR方法,对分析治疗过程的病情变化和指导治疗方案具有重要意见。
Objective To set up a real-time RT- PCR using TaqMan technology for detection and quantification of PML/RARα fusion gene transcription in acute premyeloid leukemia （APL） patients and establishment the baseline of Normal Copy Number （NCN） of the APL patients. Methods Real-time PCR was used to detect three types of PML/RARα fusion gene transcripts. Thirty-two bone marrow samples from 32 APL patients were analyzed. Results The sensitivity of real-time quantitative RT-PCR was 101copies/ul. The coeffieiency of interassay and intraassay was below 5%. Among 32 APL patients positive for PML/RARα fusion gene, 21 were found with PMI./RARct fusion gene L-type isoform, while type 11 with S-type isoform. There was no significant difference of normalized copy number （NCN） in the two groups and clinical significance. Conclusions High sensitive and specific real-time quantitative RT-PCR was successfully established for detection of PML/ RARα fusion gene in APL patients and there is no difference in the prognosis of APL patients with two isoforms.
出处
《中国热带医学》
CAS
2009年第2期260-262,共3页
China Tropical Medicine
基金
深圳市卫生局科技重大项目基金(200639)
