骨髓涂片双色荧光原位杂交检测急性早幼粒细胞白血病的PML/RARα基因重排  

Detection of PML/RARα Gene Rearrangement in Suspected Acute Promyelocytic Leukemia Patients Using Dual color Fluorescence in situ Hybridization on Bone Marrow Smears

作　　者：朱永林[1] 吴亚芳[1] 潘金兰[1] 薛永权[1] 

机构地区：[1]苏州大学附属第一医院,江苏省血液研究所苏州215006

出　　处：《中国实验血液学杂志》2004年第6期757-760,共4页Journal of Experimental Hematology

摘　　要：为了探讨双色荧光原位杂交 (D FISH)技术在骨髓涂片 (BMS)上检测PML/RARα基因重排对急性早幼粒细胞白血病 (APL)的诊断价值 ,采用光谱绿 (SpectrumGreen)和光谱桔红 (SpectrumOrange)分别标记PML和RARα两种基因探针对 2 7例临床拟诊为APL的患者进行BMS D FISH检测 ,并与常规细胞遗传学 (CCG)、逆转录 聚合酶链反应 (RT PCR)检测结果作比较。结果表明 :18例初诊患者中 ,CCG检出t( 15 ;17)者 14例 ,他们的BMS D FISH和RT PCR均证实有PML/RARα基因重排 ,从而肯定了对APL的诊断 ;而CCG未检见t( 15 ;17)易位的 4例中有 1例显示阳性的BMS D FISH和RT PCR结果 ,证实该例有隐匿易位存在 ,其余 3例的两种检测均是阴性 ,因而他们被重新诊断为AML而不是APL ;9例缓解患者中 ,CCG查出 1例部分缓解患者有t( 15 ;17)易位 ,其BMS D FISH和RT PCR均为阳性 ,而 8例完全缓解患者 ( 6例伴正常核型 ,2例未作CCG检测 )的BMS D FISH和RT PCR检测显示 6例阴性 ,2例阳性 ,提示该 2例患者有微小残留病 (MRD)存在。结论 :BMS D FISH是检测APLPML/RARα基因重排的敏感可靠方法 ,有助于APL确诊及其MRD检测 ,适合于CCG检测失败、伴有隐匿易位、RT PCR检测缺乏材料以及需作回顾性研究者。To explore the value of detection of PML/RARα gene rearrangement on bone marrow smears (BMS) by dual color fluorescence in situ hybridization (D FISH) for the diagnosis of acute promyelocytic leukemia(APL), the locus specific probes for PML and RARα genes labeled directly and respectively by Spectrum Green and Spectrum Orange and the D FISH technique were used to detect the PML/RARα gene rearrangement on BMS in 27 suspected APL patients. The results were compared with that of conventional cytogentics (CCG) and reverse transcriptase polymerase chain reaction (RT PCR). The results showed that out of 18 newly diagnosed patients 14 were found having t(15;17) translocation by CCG and PML/RARα gene rearrangement were confirmed by BMS D FISH and RT PCR. Thus, their APL diagnosis was determined; out of 4 patients in whom t(15;17) translocation was not detected by CCG, one had positive BMS D FISH and RT PCR results, thus, this case was considered as having a cryptic t(15;17) translocation, three had negative BMS D FISH and RT PCR results, thus, they were diagnosed as having acute myeloid leukemia rather than APL. In 9 cases with remission, one case with partial remission was found having t(15;17) translocation by CCG, and he had positive BMS D FISH and RT PCR results, the other 8 patients (6 cases with normal karyotype and 2 cases without CCG examination) displayed different BMS D FISH and RT PCR results: negative in 6 cases, but positive in 2 cases. The 2 cases were believed that they survived with minimal residual disease (MRD). It is concluded that BMS D FISH is a sensitive and reliable method for the detection of PML/RARα rearrangement. It is helpful for diagnosing APL and monitoring its MRD, and especially fit to those patients presenting a cryptic translocation or with failed cytogenetics, lacking suitable material for RT PCR, as well as needing retrospective study.

关 键 词：急性早幼粒细胞白血病 骨髓涂片 双色荧光原位杂交 PML/RARα基因重排 

分 类 号：R733.71[医药卫生—肿瘤] R730.4[医药卫生—临床医学]