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目的制备慢性髓细胞性白血病（ＣＭＬ）ｂｃｒ基因重排的ＤＮＡ探针。方法应用Ａｌｕ－ＰＣＲ和荧光原位杂交（ＦＩＳＨ） 技术，对３个酵母人工染色体（ＹＡＣ）候选克隆进行鉴定，制备ＤＮＡ探针，并经正常人外周血淋巴细胞和慢性髓细胞性 白血病患者骨髓细胞筛选。结果确定了ＹＡＣ７６５Ｅ３为非嵌合体，定位于染色体２２ｑｌｌ ｂｃｒ基因区域，而且可在Ｐｈ染 色体阳性细胞中易位至９ｑ３４。探针特异性杂交效率达９５％。结论ＦＩＳＨ技术是染色体、染色体畸变鉴定和染色体上特 殊序列定位的重要检测方法。联合应用Ａｌｕ－ＰＣＲ技术特异性扩增载体ＹＡＣ中人类基因组来源的ＤＮＡ制备探针，为慢性髓细胞白血病的诊断和监测微小残留病提供了一个新的手段。
Objective To prepare DNA probes for bcr gene rearrangement in chronic myelogenous leukemia (CML). METHODS: Three yeast artificial chromosome (YAC) candidate clones were identified by Alu-PCR and fluorescence in situ hybridization (FISH) techniques. DNA probes were prepared and used to pass the bone marrow of normal human peripheral blood lymphocytes and chronic myeloid leukemia patients. Cell screening. As a result, it was confirmed that YAC765E3 is a non-chimera, localized to the region of chromosome 22qll bcr gene, and translocated to 9q34 in Ph chromosome-positive cells. The probe-specific hybridization efficiency is 95%. Conclusions FISH is an important detection method for identification of chromosomes, chromosomal aberrations and localization of special sequences on chromosomes. The Alu-PCR technique was used to specifically amplify the human genome-derived DNA probes in the vector YAC to provide a new method for the diagnosis and monitoring of minimal residual disease in chronic myeloid leukemia.