双色双融合荧光原位杂交在成人急性淋巴细胞白血病遗传学异常检测中的信号模式及临床应用  

Signal Patterns of Dual Color Dual Fusion Fluorescence in Situ Hybridization for Detection of Genetic Abnormality in Adult Patients with ALL and Their Clinical Application

作　　者：江梅[1] 张长林[1] 辛艳红 刘淑媛[1] 李欣[1] 万腊根[1] 

机构地区：[1]南昌大学第一附属医院检验科,江西南昌330006 [2]江西省宜春市第二人民医院,江西宜春336000

出　　处：《中国实验血液学杂志》2016年第2期375-380,共6页Journal of Experimental Hematology

摘　　要：目的:研究双色双融合荧光原位杂交(dual-color-dual-fusion fluorescence in situ hybridization,DCDF-FISH)在成人急性淋巴细胞白血病(ALL)遗传学异常检测中的常见信号模式,并探讨其诊断价值及临床应用。方法:回顾性分析本院诊断的68例ALL病人临床资料,采用双色双融合荧光原位杂交技术、流式细胞技术、常规G显带技术以及逆转录-聚合酶链反应(RT-PCR)检测骨髓标本,并分析其结果的相关性,通过DCDF-FISH技术动态观察患者对治疗的反应。结果:在68例患者中DCDF-FISH检测见有16种信号模式,其中变异性信号模式14种(1R2G、2R3G、2R4G及3R3G为无BCR/ABL融合基因的异常信号模式;1R1G1F、1R1G3F、1R1G4F、1R2G1F、1R2G2F、1R2G3F、1Rn G2F(n≥3)、2R2G1F、1G4F、1R4F这些信号模式的细胞内都存在着BCR/ABL融合基因),Ph^+共17例,所有检出Ph^+的病例均为急性B淋巴细胞白血病(B-ALL)或伴髓系表达急性B淋巴细胞白血病(My^+-B-ALL),其中常规显带技术检测出Ph染色体12例,阳性率为18%(12/68),DCDF-FISH及RT-PCR检测结果一致,阳性检出率为25%(17/68)。通过DCDF-FISH技术动态观察患者化疗前后荧光模式变化显示:经过化疗药物的选择作用,其信号模式在数量上和形式上发生变化,共同特征是都存在Ph染色体。结论:DCDF-FISH法检测ALL病人的BCR/ABL融合基因敏感、可靠,分析DCDF-FISH信号模式及其动态变化对ALL病人的治疗反应性、耐药情况、疗效判定及预后有重要指导意义。Objective:To study the signal patterns of dual color dual fusion fluorescence in situ hybridization(DCDFFISH) for detection of genetic abnormality in adult acute lymphoblastic leukemia(ALL) patients and their diagnostic value and clinical application.Methods:The clinical data of 68 ALL patients confirmed in our hospital were analyzed retrospectively;The bone marrow samples were detected by DCDF-FISH,flow cytometry,conventional cytogenetics(CCG),reverse transcriptase polymerase chain reaction(RT-PCR),and the correlation of these results was compared.And the reaction of patients to treatment was dynamically observed by DCDF-FISH.Results:Sixteen signal patterns were found in DCDF-FISH,including 14 kinds of atypical signal patterns(signal patterns of 1R2 G,2R3G,2R4 G and 3R3 G as abnormal signal patterns without BCR/ABL fusion gene.Signal patterns of 1R1G1 F,1R1G3F,1R1G4 F,1R2G1F,1R2G2 F,1R2G3F,1RnG2F(n≥3),2R2G1 F,1G4F,1R4 F corresponded to t(9;22) karyotype).Ph^+ ALL patients accounted for 17.All cases with Ph chromosome or BCR/ABL positive were B-ALL or My^+- B-ALL.The Ph chromosome was detected in 12 cases(positive rate was 18%) by CCG.The positive rate was 25%(17/68) by DCDFFISH and RT-PCR.The DCDF-FISH fluorescence pattern change before and after chemotherapy of the patients showed that the quantity and form of the signal pattern was changed after chemotherapy,and the common characteristics was the Ph chromosome in patients.Conclusion:The DCDF-FISH is a sensitive and reliable method for the detection of BCR/ABL rearrangement.Analyzing the dynamical change of DCDF-FISH signal patterns has been comfirmed to have a important guiding significance in the diagnosis,and anlysis of response to therapy,drug resistance and the prognosis of ALL patients.

关 键 词：急性淋巴细胞白血病 双色双融合荧光原位杂交 遗传学异常 BCR/ABL融合基因 费城染色体 

分 类 号：R733.71[医药卫生—肿瘤]