CAJ | 학술논문

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餐检测,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ber-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ber-ablmRNA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,可榆出allo-HSCT后发生分子生物学或细胞遗传学复发的高危患者.
목적 탐토이용단천련중복서렬취합매련반응(STR-PCR)결합반전록취합매련반응(RT-PCR)정량화정성검측이기인조혈간세포이식(allo-HSCT)후환자감합체화융합기인적표체,분석기식입화미소잔류병변정황,평개기대복발적예측개치.방법 감합체공체세포감합솔채용STR-PCR결합모세관전영진행정찬검측,대융합기인전록본bcr-abl mRNA채용RT-PCR방법검측.결과 5례환자재이식후+28천균위100%공자형감합,융합기인ber-abl mRNA균위음성.단재이후적수방과정중발현5례환자재불동시간출현불은정혼합감합(MC)상태[공체세포감합(DC)솔위0～80.4%].융합기인ber-ablmRNA양성.기중1례복발후일직처우MC,최후사망.령외4례재림상간예치료후우전변위완전감합(CC),목전처우분자생물학완해상태.상술5례복발환자균재출현림상증상전발생DC하강;융합기인표체양성.결론 STR-PCR재민감범위내,기결과여RT-PCR적결과부합솔고,량충기술결합대검측allo-HSCT후공체시부식입、질병복발이급이식물항숙주병(GVHD)균유예경작용,대실시림상간예치료유중요지도개치,가유출allo-HSCT후발생분자생물학혹세포유전학복발적고위환자.
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 alIo-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 ehimerism. (DC: 0～80.4 %) at the different time points after aIIo-HSCT and bcr-abl was positive. One of them kept eontinuely the mixed chimerism in the relapse of disease, and died after one year, and the other 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.