多重巢式RT-PCR和染色体核型分析技术在急性髓系白血病中的联合应用

Combined Application of Multiplex Reverse Transcription-Polymerase Chain Reaction and Karyotype Analysis to Detection of Clonal Chromosomal Aberrations in Acute Myeloid Leukemia

背景与目的:细胞遗传学分析在白血病的诊断和预后判断中有重要价值,但常规显带不仅耗时,且难以获得良好的分裂相;而聚合酶链反应具有灵敏、快速的特点。本研究探讨联合应用多重巢式RT-PCR和染色体核型分析,对急性髓系白血病(acute myeloid leukemia,AML)中融合基因的表达及其在各亚型的分布和克隆性染色体异常的检出情况。方法:采用多重巢式RT-PCR技术对60例AML病例进行检测,其中37例同时进行染色体R或G显带。结果:60例AML患者中检出融合基因28例(46.7%),包括AML1/ETO、PML/RARα、CBFβ/MYH11、MLL基因异常(包括MLL/AF6、MLL/AF9、MLL/AF10、MLL/MLL)、DEK/CAN、TEL/PDGFR、AML1/MDS1(EVI-1)。同时进行染色体R或G显带的37例病例中有30例可供分析,其中14例(46.7%)检出染色体结构和数目异常;联合多重RT-PCR可使AML克隆性染色体异常检出率增至59.5%(22/37)。结论:联合多重巢式RT-PCR和染色体核型分析技术可以提高克隆性染色体异常的检出率。

BACKGROUND ＆ OBJECTIVE： Cytogenetic analysis plays a critical role in the diagnosis and prognosis evaluation of leukemia, but karyotype analysis is time-consuming and difficult to yield sufficient metaphases; while polymerase chain reaction （PCR） is sensitive and efficient. This study was to investigate combined application of multiplex reverse transcription-polymerase chain reaction （RT-PCR） and karyotype analysis to the detection of clonal chromosomal aberrations in acute myeloid leukemia （AML） , and explore the expression and distribution of fusion genes among the subtypes of AML. METHODS： Sixty AML patients were examined by multiplex RT-PCR. Cytogenetic data were obtained from 37 of them by R or G banding techniques. RESULTS： Fusion genes, including AML1 /ETO, PML/RARα, CBFβ/MYH11, MLL gene rearrangements （that is, MLL/AF6, MLL/AF9, MLL/AF10, and MLL/MLL） , DEK/CAN, TEL/PDGFR, and AML1 /MDS1 （EVI-1） , were detected in 28 （46.7% ） patients by multiplex RT-PCR. In the 37 patients who received karyotype analysis, data were available in 30 patients and cytogenetic aberrations were detected in only 14 （ 46.7%） of them. The detection rate of clonal chromosomal aberrations was enhanced to 59.5% by combined application of multiplex RT-PCR and karyotype analysis. CONCLUSION： Multiplex RT-PCR combined with karyotype analysis can improve the detection rate of clonal chromosomal aberrations in AML.