多重巢式反转录-聚合酶链反应检测白血病相关融合基因及其临床意义

Detection of the fusion genes by multiplex RT-PCR and its clinical significances in leukemia

目的检测白血病相关29种染色体结构畸变形成的融合基因并对其在白血病MICM分型、危险度分组和微小残留病（MRD）检测中的临床价值进行分析。方法采用多重巢式反转录-聚合酶链反应（RT—PCR）方法对141例白血病患者进行29种染色体结构畸变形成的融合基因进行分析，并与骨髓染色体检测结果进行对比分析。结果141例白血病患者的骨髓或外周血标本中，66例（46．8％）阳性，具有13种染色体结构畸变产生的融合基因，包括PML／RARα、bcr-abl p210、bcr—abl p190、AML1／ETO、TEL／AML1，E2A／PBX1、MLL／AFX、MLL／AF6、MLL／AF9、dupMLL、MLL／ENL、CBFβ／MYH11、TLS／ERG等。在78例ALL和57例AML标本中，分别有27例（47．4％）和33例（42．3％）为阳性，染色体结构畸变产生的融合基因分别为7种和6种。结论采用多重巢式RT—PCR方法，可快速分析29种染色体结构畸变产生的融合基因，可同时筛选出29种急性白血病的常见染色体易位，帮助临床诊断、疾病危险度分组、预后判断和治疗后微量残留病的检测。

Objective To analyze the fusion genes derived from 29 types of chromosome structural aberrations in leukemia patients, and the significances on the MICM typing, risk grouping, and minimal residual disease （MRD） monitoring of leukemia. Methods The bone marrow or blood samples from 141 leukemia patients were analyzed with a novel muhiplex nested RT-PCR. In addition, chromosomal karyotypes were investigated in some patients. Results Of the 141 leukemic samples, 66 （46.8 %） carried 13 types of fusion genes including PML/RARα, hcr/ab1 p210, bcr/abl p190, AML1/ETO, TEL/AML1, E2A/PBX1, MLL/AFX, MLL/AF6, MLL/AF9,dupMLL, MLL/ENL, CBFβ/MYH11 and TLS/ERG. Fusion genes were positive in 27 of 57 ALL patients （47.4 %）, and 33 of 78 AML patients （42.3 %）, respectively. In these ALL or AML patients, 7 or 6 chromosome structural aberrations were found. Conclusion This multiplex nested RT-PCR reaction could screen 29 types of chromosome structural aberrations at the same time. It may be helpful for the diagnosis, risk grouping, prognosis evaluation and the detection of minimal residual diseases after chemotherapy and bone marrow transplantation in these leukemia patients.