间期双色双融合荧光原位杂交监测慢性髓系白血病异基因造血干细胞.docVIP

间期双色双融合荧光原位杂交监测慢性髓系白血病异基因造血干细胞 　作者：钱思轩，李建勇，张闰，洪鸣，仇海荣，李丽，徐卫，盛瑞兰，吴汉新 【摘要】 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。 【关键词】 荧光原位杂交 　　Monitoring of bcr/abl Fusion Gene by Interphase-Dual-Color and Dual-Fusion Fluorescence in situ Hybridization in CML after Allo-HSCT 　　Abstract This study was aimed to investigate the sensitivity and clinical application value of interphase-dual-color and dual-fusion fluorescence in situ hybridization (DD-FISH). The minimal residual disease (MRD) in 19 patients with chronic myelogenous leukemia (CML) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) was detected by DD-FISH，and the detected results were compared with those of conventional cytogenetics (CC) and reverse transcription-polymerase chain reaction (RT-PCR).The samples were collected from bone marrow or peripheral blood or smears of bone marrow. The results indicated that 14 out of 19 patients achieved and maintained continuous complete molecular remission after transplantation. In these patients，CC assay displayed normal donor karyotype，result of RT-PCR was negative，complete donor chimerism was detected after 2 months of transplantation，result of DD-FISH was negative，average time of the follow-up survey was 11.25 months，MRD did not increase. Results of CC and RT-PCR in 1 patient showed negative，while FISH of sex chromosome showed mixed chimerism，result of DD-FISH was positive，MRD did not increase，no therapy was given for this patiant，clinical state