应用性染色体双色间期荧光原位杂交检测异性间造血干细胞移植后嵌合状态

Application of Dual-color Fluorescence in Situ Hybridization in Study on Chimeric Status after Sex-mismatched Allogeneic Hematopoietic Stem Cell Transpiantation

目的 应用性染色体双色间期荧光原位杂交技术检测异性间造血干细胞移植后,患者的嵌合状态,并对其应用价值进行分析.方法 对2006年1月～2007年12月在天津血液病医院行异性间allo-HSCT的32例患者,常规染色体核型分析采用不加任何刺激剂的骨髓24 h短期培养法制备染色体,火焰法滴片,R显带.应用X/Y性染色体双色着丝粒DNA探针对骨髓间期细胞行荧光原位杂交,检测移植后患者的嵌合状态.结果 32例患者染色体核型分析115例次,其中111例次为正常供者核型,1例次受者核型,3例次分析失败,未能检测到混合核型.双色性染色体间期FISH检测163例次,129例次为完全嵌合,34例次为混合嵌合,检测成功率为100%.移植后2～3 w 32.2%患者为混合状态,移植后1 mo 76.7%患者为完全嵌合,移植后半年94.1%患者为完全嵌合.结论 性染色体双色间期荧光原位杂交能够简便、快速、灵敏地检测出异性间allo-HSCT患者移植后的嵌合状态.

Objective Application of dual-color fluorescence in situ hybridization in study on chimeric status after sex-mismatched allogeneic hematopoietic stem cell transplantation,and evaluate the value of dual-color interphase fluorescence in situ hybridization （I-FISH） in detection of chimerism. Methods 32 patients with blood diseases received allogeneic hematopoietic stem cell grafts from sex-mismatched donors. Bone marrow cells were performed by 24 h unstimulated culture, R-banding and karyotype by conventional cytogenetic analysis （CCA）. Chimerism were investigated by I-FISH with dual-color sex chromosome centromere DNA probe in interphase cells of bone marrow. Results CCA showed karyotype of donors in 111 and karyotype of recipient in 1. Entire chimerism sex chromosome were identified by I-FISH in 163,mixed chimerism in 34. Chimerism of 32.2% patients in two or three weeks after allogeneic hematopoietic stem cell transplantation was mixed. The proportion of entire chimeric patients was 76.7% and 94.1% in one month and six month after allo- HSCT. Conclusion I-FISH could be used as a convenient,rapid and sensitive method to detect chimerism after sex-mismatched allogenelc hematopoietic stem cell transplantation.