实时定量PCR技术检测BCR-ABL水平在异基因造血干细胞移植治疗慢性粒细胞白血病中的应用

目的探讨实时定量PCR（qRT-PCR）技术检测慢性粒细胞白血病（CML）患者异基因造血干细胞移植（allo．HSCT）治疗后BCR．ABL水平对改善患者预后的临床价值。方法研究纳入河北联合大学附属医院血液科2005年5月至2013年5月接受allo-HSCT治疗的CML患者15例，其中慢性期患者8例，加速期患者3例，急变期患者4例。分别在治疗前及治疗后1，2，3，6，12个月留取骨髓血标本，采用基于TaqMan探针的qRT-PCR技术检测BCR-ABL水平。根据BCR-ABL水平的动态变化，评估患者复发程度，对高复发风险的患者给予早期干预性治疗。两组问均数比较采用t检验。结果慢性期及进展期（加速期＋急变期）患者在治疗前平均BCR—ABL水平分别为（10．120±6．035）％及（43．750±14．173）％，差异具有统计学意义（t=-2．289，P〈0．05）。治疗后第1，2，3，6，12个月，慢性期和进展期患者平均BCRABL水平相比，差异均无统计学意义（t=-1．936，-2．003，-0．687，-1．613和-1．171，均P〉0．05）。治疗后第12个月，8例慢性期患者的BCR-ABL水平均为0，7例进展期患者中有5例为0。2例慢性期患者和5例进展期患者接受早期干预性治疗，除1例急变期患者在治疗后12个月复发，行再次移植无效而死亡外，其余6例患者均达完全分子生物学缓解，至随访截止时均无病存活。结论qRT-PCR是评估CML患者治疗后复发情况的良好方法，通过BCR-ABL水平的动态监测，可筛选出高复发风险的患者，配合早期干预性治疗措施可以提高患者无病生存率。

Clinical significance of detection of BCR.ABL level by qRT-PCR in patients with chronic myelocytic leukemia after allogeneic hematopoietic stem cell transplantation

Objective In order to reduce the relapse rate and improve disease-free survival of chronic myelocytic leukemia （CML） patients after allogeneic hematopoietic stem cell transplantation （allo-HSCT） , the timely, effective and reasonable intervenetion measures were implemented to the CML patients with high recurrence risk after allo-HSCT screened out according to the BCR-ABL mRNA dynamic monitoreing by real time quantitative PCR （qRT-PCR）. Methods Eight CML chronic phase （CML-CP） cases and 7 CML advanced phase （CML-AP） cases after allo-HSCT were retrospectively analyzed from May 2005 to May 2013 in the Hebei United University Affiliated Hospital, bone marrow samples acquired respectively before transplantation or in 1,2, 3, 6 and 12 months after transplantation were detected by qRT- PCR technique with TaqMan probe. According to the BCR- ABL level changes, the early interventional therapies were implemented to the patients with high recurrence risk, for example, taking imatinib or decrement immunosuppressive agents or combination of both methods. The differences between two groups were determined by t test. Results Before allo-HSCT, the BCR-ABL level was （ 10. 120 ± 6. 035 ） % and （43. 750 ± 14. 173 ） % in CML- CP and CML- AP cases, respectively （ t = -2. 289, P 〈0. 05）. After treatment, there were no significant difference in BCR-ABL level between CML-CP and CML-AP patients at each time point. At the month 12th after allo-HSCT, the BCR-ABL levels of 8 CML-CP and 2 CML-AP patients were zero. Two CML-CP and 5 CML-AP patients were received interventional treatment, 1 CML-AP patient died from recurrence and others got molecular remission. Conclusions qRT-PCR is a sensitive, reliable method for evaluating recurrence of the CML patients after allo-HSCT. The early intervenetion measures implemented to the CML patients with high recurrence risk after allo-HSCT by dynamic monitoring of BCR-ABL level can reduce the recurrence rate and improve the disease-free survival rate after allo-HSCT.