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本研究探讨bcr/abl双色双融合荧光原位杂交(DD-FISH)的敏感性及临床应用价值。对19例慢性髓细胞白血病(CML)异基因造血干细胞移植(allo-HSCT)后微小残留病灶(MRD)用DD-FISH进行监测,同时与常规细胞遗传学(CC)、逆转录-聚合酶链反应(RT-PCR)结果相比较。样本取自骨髓,少数来源于骨髓片或外周血。结果表明:14例CML患者在Allo-HSCT后CC显示持续正常供者核型,RT-PCR转为阴性,移植2月后均为完全供者嵌合(DC),DD-FISH检测结果持续为阴性,平均随访11.25月,MRD无增加。1例CC及RT-PCR阴性,而性染色体FISH为混合嵌合,DD-FISH阳性,监测无MRD增加,临床未治疗,疾病稳定。3例骨髓复发患者的DD-FISH及性染色体FISH均提示MRD明显增加,RT-PCR转为阳性,却只有1例CC异常,供者淋巴细胞输注(DLI)及甲磺酸伊马替尼治疗后均为DC,DD-FISH阴性,RT-PCR阴性。1例骨活检证实髓外复发患者骨髓或外周血样本的DD-FISH、CC及PCR均阴性,供受者完全嵌合。结论:间期双色双融合FISH可应用于CML异基因造血干细胞移植后MRD的监测,其操作简易快速,灵敏度高,且骨髓或外周血均可采用。动态监测能及时发现扩大的白血病细胞克隆。
This study was to explore the sensitivity and clinical value of bcr / abl dual-color double-fusion fluorescence in situ hybridization (DD-FISH). DD-FISH was used to monitor the minimal residual disease (MRD) in 19 patients with chronic myelogenous leukemia (CML) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Meanwhile, it was compared with conventional cytogenetics (CC), reverse transcription polymerase chain Reaction (RT-PCR) results were compared. Samples were taken from the bone marrow and a few were from bone marrow or peripheral blood. The results showed that in 14 CML patients, CC showed continuous normal karyotype after Allo-HSCT, RT-PCR turned negative, and all were completely donor chimerism (DC) after 2 months of transplantation. The results of DD-FISH continued to be Negative, average follow-up 11.25 months, MRD no increase. One case was negative for CC and RT-PCR while the other was chromosomal FISH for mixed chimerism. DD-FISH was positive. No MRD was detected in the study. The disease was clinically untreated and the disease was stable. Both DD-FISH and sex chromosome FISH in 3 patients with bone marrow recurrence suggested that MRD increased significantly and RT-PCR turned positive, only 1 patient had CC abnormality, donor lymphocyte infusion (DLI) and imatinib mesylate After treatment were DC, DD-FISH negative, RT-PCR negative. DD-FISH, CC and PCR were negative in 1 bone marrow biopsy or bone marrow biopsy in patients with extramedullary recurrence, and the recipients were completely chimeric. Conclusion: Interphase bi-color double-fusion FISH can be used to monitor MRD after CML allogeneic hematopoietic stem cell transplantation. It is simple, rapid and sensitive, and can be used in bone marrow or peripheral blood. Dynamic monitoring can detect enlarged leukemia cell clones in time.