实时定量聚合酶链反应监测慢性粒细胞白血病患者接受伊马替尼治疗期间bcr-abl融合基因拷贝数的变化

 目的　应用实时定量聚合酶链反应（RQ-PCR）技术动态监测接受伊马替尼(IM)治疗的慢性粒细胞白血病（CML）患者bcr-abl融合基因拷贝数的变化，探讨RQ-PCR技术在微小残留病（MRD）检测以及预测复发方面的应用。方法　应用RQ-PCR技术动态监测106例接受IM治疗的CML患者bcr-abl融合基因拷贝数，bcr-abl融合基因定量结果以校正比值（NQ）表示，NQ＝bcr-abl拷贝数／abl拷贝数。结果　IM治疗前患者的NQ值与其疾病进展及Ph+细胞数量均显著相关（r＝0.9824，r＝0.9346）。使用IM治疗的106例患者，62例在治疗12个月内NQ值迅速下降并长时间维持在较低水平，其中仅2例复发；8例在治疗后NQ值略有下降但随后又马上升高，其中7例在NQ值升高后的5～9个月内复发；31例治疗后NQ值未见明显下降且仍>0.1，其中11例获得过短暂的形态学缓解，随后迅速复发，7例治疗无效或疾病进展；另有5例虽然NQ值波动较大，且无规律性，但治疗后一直处于形态学缓解。结论　RQ-PCR方法准确、可靠、敏感度高，在监测CML患者的MRD、判断疗效以及预测复发等方面具有重要的临床应用价值。

Expression patterns of bcr-abl measured by real-time quantitative PCR in patients with chronic myeloid leukemia during treatment with imatinib mesylate

Objective To monitor the expression patterns of bcr-abl in chronic myeloid leukemia (CML) patients during treatment with imatinib mesylate and evaluate the detection of MRD by RQ-PCR method. Methods The ABI Prism 7500 Sequence Detection System using Taqman fluorogenic probes was used to quantify target gene. bcr-abl mRNA was detected by RQ-PCR in 106 CML patients. The normalized quotient (NQ) of bcr-abl mRNA was calculated as followings: NQ=bcr-abl mRNA copy numbers/abl mRNA copy numbers. Results The NQ of BCR-ABL mRNA was well correlated with the progression of disease and the number of Ph+ cell (r =0.9824 and 0.9346, respectively). The NQ was decreased rapidly in 62 patients and kept in low level for a long time, and only 2 of them were relapsed. For 8 patients, after treatment the NQwere decreased initially and increased sharply, 7 of them were relapsed after 5-9 months. After treatment the NQ of 31 patients were still>0.1, 11 patients were relapsed after a short remission and 7 were ineffective or progression. Out of 5 patients whose NQ were fluctuated and had little regularity, but all of them had a continuing remission. Conclusion RQ-PCR is a more sensitive technique in the detection of bcr-abl fusion gene.It is an important method to monitor the tumor cell during the treatment with imatinib mesylate in CML patients.