Alu-PCR联合荧光原位杂交技术鉴定bcr基因相关YAC克隆  

Identification of YAC clone related to bcr gene by Alu-PCR and fluorescence in situ hybridization

作　　者：刘晓力[1] 郑维扬[1] 刘孟珉[2] 宋兰林[1] 杜庆锋[1] 陈赛娟[2] 周淑芸[1] 

机构地区：[1]第一军医大学南方医院血液科,广东广州510515 [2]上海第二医科大学瑞金医院血液学研究所,上海200025

出　　处：《第一军医大学学报》2001年第5期321-323,F002,共4页Journal of First Military Medical University

基　　金：国家自然科学基金!(39770831);广东省自然科学基金!(970833)

摘　　要：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 probe detecting bcr gene rearrangement in chronic myeloid leukemia (CML) by fluorescence in situ hybridization (FISH). Methods Three clones of yeast artificial chromosomes (YAC) related to bcr gene were examined by FISH in combination with Alu-PCR to identify metaphases peripheral lymphocytes from normal individuals and metaphases bone marrow cells from CML patients with chromosome t (9; 22). Results One of 3 YAC, 765E3 DNA probe was verified without evidence of chimerism at normal metaphases by FISH, which can translocate to chromosome 9 q34 from 22 qll in Ph (+) cells. Its efficiency for specific hybridization was 95%. Conclusion FISH is a versatile technique for identifying chromosomes, detecting chromosomal abnormalities or determining the chromosomal localization of specific sequence. It provides a new tool for the diagnosis and monitoring of minimal residual disease in CML.

关 键 词：聚合酶链反应 荧光原位杂交 酵母人工染色体 BCR基因 慢性髓细胞白血病 YAC DNA探针 

分 类 号：R733.72[医药卫生—肿瘤] Q754[医药卫生—临床医学]