应用三色双融合探针检测BCR-ABL融合基因及ASS1基因缺失  

Detection of BCR/ABL Fusion Gene and ASS1 Gene Deletion by Using Tricolor Dual-fusion Probe

作　　者：张朕豪 王艳芳[1] 王淼[2] 董菲[1] 万伟[1] 克晓燕[1] 景红梅[1] ZHANG Zhen-Hao;WANG Yan-Fang;WANG Miao;DONG Fei;WAN Wei;KE Xiao-Yan;JING Hong-Mei(Department of Hematology,Peking University Third Hospital,Beijing 100191,China;Department of Pediatrics,Peking University People′s Hospital,Beijing 100044,China)

机构地区：[1]北京大学第三医院血液科,北京100191 [2]北京大学人民医院儿科,北京100044

出　　处：《中国实验血液学杂志》2020年第4期1115-1122,共8页Journal of Experimental Hematology

基　　金：国家自然科学基金(81700192)。

摘　　要：目的:分析BCR/ABL/ASS1三色双融合探针在CML和B-ALL所出现的各种异常信号模式的意义,并探讨其在检测BCR/ABL融合基因及ASS1基因缺失中的应用价值。方法:分别对50例初诊CML患者和50例初诊B-ALL患者采用BCR/ABL/ASS1三色双融合探针进行荧光原位杂交(FISH)检测,同时对所有病例应用24 h短期培养法R显带后进行染色体核型分析。结果:50例CML患者中,49例Ph^+,余1例可见5个正常中期核型;通过FISH发现,所有患者(100%)均存在BCR/ABL融合基因,阳性信号特征分别为1R1G2B2F 39例(78%)、2R1G2B1F 2例(4%)、1R1G1B1F 6例(12%)、同时伴有1R1G2B2F和1R1G2B3F 2例(4%)、2R2G2B1F 1例(2%)。伴有ASS1基因缺失(1R1G1B1F)的6例患者染色体核型均为单纯t(9;22)易位,无其他异常。50例B-ALL患者中,Ph^+13例,数目畸变及非t(9;22)的结构畸变16例,正常核型20例,无分裂相1例;经FISH检出16例(32%)具有BCR/ABL融合基因,分别为1R1G2B2F 13例(26%)、同时伴有1R1G2B2F和1R1G2B3F 1例(2%)、2R1G1B1F 1例(2%)、1R1G3B3F 1例(2%),FISH额外检出的3例BCR/ABL融合基因阳性包括1例伴有ASS1基因缺失(2R1G1B1F)、1例经典型t(9;22)易位(1R1G2B2F)和1例BCR/ABL融合基因及ASS1基因拷贝数的增加(1R1G3B3F)。结论:应用三色双融合FISH探针检测BCR/ABL融合基因及ASS1基因缺失,具有简单、快速、灵敏、稳定的特点,能够检测多种形式的分子融合,可很好地避免D-FISH探针及ES-FISH探针因信号随机重叠导致的假阳性结果。该检测方法不但可以直接观察有无ASS1基因的缺失,而且还能提高初诊BCR/ABL融合基因阳性结果的可靠性及复诊监测微小残留病结果的准确性。Objective:To analyze the significance of various abnormal signal patterns appreared in CML and B-ALL patients by using BCR/ABL/ASS1 tricolor dual-fusion probe,and to explore its application value in detecting BCR/ABL fusion gene and ASS1 gene deletion.Methods:50 newly diagnosed CML patients and 50 newly diagnosed B-ALL patients were detected by fluorescence in situ hybridization(FISH)with BCR/ABL/ASS1 tricolor dual-fusion probe.Meanwhile,karyotype analysis was performed on all the patients using the 24 hours short-term culture and R-banding.Results:Among the 50 CML patients,Ph^+was found in 49 cases,5 normal interphase karyotype was observed in 1 case.FISH detection showed that BCR/ABL fusion gene existed in all patients(100%),while the positive signal pathway showed that 1R1G2B2F was observed in 39 cases(78%),2R1G2B1F in 2 cases(4%)and 1R1G2B1F in 6 cases(12%),simultaneous existence of 1R1G1B1F and 1R1G2B3F in 1 case(2%),2R1G1B1F in 1 case(2%)1R1G3B3F in 1 case(2%).FISH detection also showed that the karyotype of 6 case at ASS1 gene deletion(1R1G1B1F)all were simple t(9;22)translocation,and other abnormalities not were observed.Among 50 cases of B-ALL,Ph^+was found in 13 cases,the numerical aberration and structural aberration of non t(9;22)in 16 cases,normal karyotype in 20 cases,absence of mitotic phase in 1 case.FISH detection showed that 16 cases(32%)had BCR/ABL fusion gene including 13 cases(26%)of 1R1G2B2F,1 case(2%)of stimultaneous exitance of 1R1G2B2F and 1R1G3B3F 1 case(2%)of 2R1G1B1F,1 case(2%)of 1R1G3B2F.FISH detection also showed that 3 cases had BCR/ABL fusion gene,including 1 case with ASS1 gene deletion(2R1G1B1F),1 case with classical t(9;22)translocation(1R1G2B2F)and 1 case with BCR/ABL fusion gene and increase of ASS1 gene copy(1R1G3B3F).Conclusion:Tricolor dual-fusion FISH probe for detecting BCR/ABL fusion gene and ASS1 gene deletion is simple,rapid,sensitive and stable.It can detect various forms of molecular fusion and avoid the false positive results due to coincidental overlap of signals g

关 键 词：三色双融合探针 BCR/ABL融合基因 ASS1基因 荧光原位杂交 

分 类 号：R733.7[医药卫生—肿瘤]