STR—PCR结合RT—PCR检测慢性粒细胞白血病造血干细胞移植后复发时嵌合体和融合基因  

Monitoring of chimerism and fusion gene by STR-PCR and RT-PCR in relapse chronic myeloid leukemia patients after hematopoietic stem cell transplantation

作　　者：孙敬芬[1] 韩晓苹[1] 金红实[1] 高春记[1] 于力[1] 

机构地区：[1]解放军总医院血液科,北京100853

出　　处：《白血病．淋巴瘤》2009年第7期392-395,共4页Journal of Leukemia & Lymphoma

基　　金：基金项目：首都医学发展科研基金重点支持项目（2007-2040）

摘　　要：目的探讨利用短串联重复序列聚合酶链反应（STR—PCR）结合反转录聚合酶链反应（RT—PCR）定量和定性检测异基因造血干细胞移植（allo—HSCT）后患者嵌合体和融合基因的表达，分析其植入和微小残留病变情况，评价其对复发的预测价值。方法嵌合体供体细胞嵌合率采用STR—PCR结合毛细管电泳进行定量检测，对融合基因转录本bcr—abl mRNA采用RT-PCR方法检测。结果5例患者在移植后＋28天均为100％供者型嵌合，融合基因bcr-abl mRNA均为阴性。但在以后的随访过程中发现5例患者在不同时间出现不稳定混合嵌合（MC）状态[供体细胞嵌合（DC）率为0．80．4％]，融合基因bcr-abl mRNA阳性。其中1例复发后一直处于MC，最后死亡。另外4例在临床干预治疗后又转变为完全嵌合（CC），目前处于分子生物学缓解状态。上述5例复发患者均在出现临床症状前发生DC下降；融合基因表达阳性。结论STR—PCR在敏感范围内，其结果与RT-PCR的结果符合率高，两种技术结合对检测allo—HSCT后供体是否植入、疾病复发以及移植物抗宿主病（GVHD）均有预警作用，对实施临床干预治疗有重要指导价值，可检出allo—HSCT后发生分子生物学或细胞遗传学复发的高危患者。Objective To investigate the value of the multiple short tandem .repeat （STR） amplification by fluorescence labeling polymerase chain reaction （PCR） combined with fusion gene bcr-abl mRNA expression for quantitative determination of chimerism and qualitative detection of bcr-abl transcripts, and to evaluate the status of engraftment and predict the outcome of allogeneic hematopoietic stem cell transplantation （allo-HSCT）. Methods 5 relapse patients with CML after allo-HSCT were dynamically investigated. Quantitative analysis of donor chimerism was performed by multiplex PCR amplification of STR markers and capillary electrophoresis with fluorescence detection, qualitative detection of bcr-abl transcripts was performed by RT-PCR. Results The donors alleles appeared in all of 5 patients on day 28 post transplant, and bcr-abl expression was negative. But 5 patients had unstable mixed chimerism. （DC： 0- 80.4 %） at the different time points after allo-HSCT and bcr-abl was positive. One of them kept continuely the mixed chimerism in the relapse of disease, and died after one year, and the other 4 changed from MC to CC by intervention of clinical treatment. Reduction of donor chimerism were detected prior to the occurrence of graft rejection and disease relapse, while bcr-abl gene expression was positive. Conclusion The results of STR-PCR in the range of its sensitivity fully correspond with bcr-abl tests in patients with CML. The combination of STR-PCR with RT-PCR provides a highly sensitive and valuable tool for engraftment evaluation, graft rejection, relapse and predicting GVHD. Furthermore it can provide a basis for early intervention of clinical treatment, and can identify these patients at high risk with molecular or cytogenetic relapse after allo-HSCT.

关 键 词：造血干细胞移植 串联重复序列 嵌合体 基因融合 

分 类 号：R733.7[医药卫生—肿瘤]