Bock Oliver, Reising Dunja, Kreipe Hans
Institute of Pathology, Medizinische Hochschule Hanover, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany.
Diagn Mol Pathol. 2003 Sep;12(3):119-23. doi: 10.1097/00019606-200309000-00001.
Diagnosis of chronic myeloid leukemia and acute lymphoblastic leukemia requires the investigation of the Philadelphia chromosome translocation t(9;22) or the molecular detection of BCR-ABL fusion transcripts. Determination of the type of fusion transcript is crucial for quantitative molecular monitoring the course of the disease during treatment. Histopathologists, who usually use formalin-fixed tissues, may be confronted with the need to investigate the BCR-ABL rearrangement when evaluating tumor forming infiltrates and bone marrow trephines from patients presenting with chronic myeloproliferative disorders. Therefore, we have established a one-tube multiplex RT-PCR for the detection of common BCR-ABL fusion transcripts (b2a2, b3a2, e1a2) in routinely processed tissues and bone marrow trephines with respect to the inevitable fragmentation of ribonucleic acids in these specimens. RT-PCR products allow distinct and unequivocal differentiation of the underlying fusion in either the Major- or minor-breakpoint cluster region. Detection of BCR-ABL fusion transcripts by multiplex RT-PCR in routinely processed and fixed tissues is a time- and cost-sparing tool for definite diagnosis of typical chronic myeloid leukemia and Philadelphia chromosome positive acute lymphoblastic leukemia.
慢性髓性白血病和急性淋巴细胞白血病的诊断需要检测费城染色体易位t(9;22)或BCR-ABL融合转录本的分子检测。确定融合转录本的类型对于在治疗期间对疾病进程进行定量分子监测至关重要。组织病理学家通常使用福尔马林固定的组织，在评估患有慢性骨髓增殖性疾病患者的肿瘤形成浸润和骨髓活检组织时，可能需要检测BCR-ABL重排。因此，考虑到这些标本中核糖核酸不可避免的片段化，我们建立了一种单管多重RT-PCR方法，用于检测常规处理的组织和骨髓活检组织中常见的BCR-ABL融合转录本（b2a2、b3a2、e1a2）。RT-PCR产物能够明确区分主要或次要断点簇区域中的潜在融合。通过多重RT-PCR在常规处理和固定的组织中检测BCR-ABL融合转录本，是明确诊断典型慢性髓性白血病和费城染色体阳性急性淋巴细胞白血病的一种节省时间和成本的工具。
