联合应用细胞遗传学、巢式RTPCR及FISH技术检测慢性髓系白血病.doc

联合应用细胞遗传学、巢式RTPCR及FISH技术检测慢性髓系白血病 　 作者：王慧萍　李国霞　 乔振华　 王宏伟 论文代写 【摘要】 　　本 研究 探讨常规细胞遗传学(conventional cytogenetics, CC)、巢式逆转录聚合酶链反应(nestedreverse transcriptase polymerase chain reaction, nestedRTPCR)及双色双融合荧光原位杂交(dualcolor and dualfusion fluorescence in situ hybridization, DFISH) 三种技术监测慢性髓系白血病(chronic myeloid leukemia，CML)患者造血干细胞移植 治疗 过程中肿瘤负荷的灵敏度和特异性。联合 应用 CC、巢式RTPCR 和DFISH三种技术对7例CML患者非清髓性异基因干细胞移植治疗前后的肿瘤负荷水平进行检测。检测结果显示： 7例CML患者治疗前后的40份骨髓标本中，有29份标本检出不同比率的Ph染色体；3份因细胞数少CC 分析 失败；36份标本RTPCR检测结果为阳性。病例1移植后12、18、26及38个月的4份标本Ph染色体及RTPCR结果均为阴性。病例1移植后9、10个月、病例2移植后15个月、病例3移植后12个月的4份Ph(-)bcr/abl(+)标本经FISH检测，分别检出5.4%、 0%、 16.5%及1.5%的bcr/abl(+)细胞。病例5移植后20、60天、病例7移植后40天的3份标本因细胞数少而CC核型分析失败，对其行FISH检测，结果bcr/abl(+)细胞检出率分别为55.0%、27.5%和73.5%。病例1移植后12个月Ph(-)bcr/abl(-)的标本行FISH检测，结果bcr/abl(+)细胞检出率为0%。结论： CC可作为监测CML患者治疗过程中肿瘤负荷水平的基本手段。在移植后早期细胞数太少而无法进行CC检测，以及治疗病人体内肿瘤负荷降低到CC不能检出而RTPCR仍为阳性时，借助FISH准确检测体内肿瘤负荷，以监测其动态变化。FISH检测bcr/abl转阴的病人需靠灵敏度更高的RTPCR监测核定。 【关键词】 慢性髓系白血病；细胞遗传学；聚合酶链反应；荧光原位杂交 　　Detection of Tumor Load in Chronic Myeloid Leukemia During Treatment with Transplantation by Conventional Cytogenetics, NestedRTPCR and FISH Abstract This study was purposed to investigate the sensitivity and specificity of conventional cytogenetics (CC),nestedreverse transcriptase polymerase chain reaction (nestedRTPCR) and dualcolor / dualfusion fluorescence in situ hybridization (DFISH) technique in monitoring the tumor load of chronic myeloid leukemia (CML) during treatment with transplantation. CC, nestedRTPCR and interphase DFISH were simultaneously carried out to detect the tumor load of 7 CML patients during treatment with nonmyeloablative allogentic stem cell transplantation(alloNSCT).40 specimens from 7 CML patients before and after alloNSCT were analyzed. The results showed that 29 specimens were Ph(+) with different positive ratio and 3 specimens with lower cells were not analyzed by CC. 36 specimens were bcr/abl mRNA (+) by RTPCR. 4 specimens from case 1 at 12