Detection of minimal residual disease in patients with AML1/ETO-associated acute myeloid leukemia using a novel quantitative reverse transcription polymerase chain reaction assay
Detection of minimal residual disease in patients with AML1/ETO-associated acute myeloid leukemia using a novel quantitative reverse transcription polymerase chain reaction assay
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DOI：
10.1038/sj.leu.2401128
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发表时间：
1998-09-01
期刊：
影响因子：
11.4
通讯作者：
Caligiuri, MA
中科院分区：
文献类型：
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作者：
Marcucci, G;Livak, KJ;Caligiuri, MA
用逆转录聚合酶链式反应(RT-PCR)检测长期完全缓解(CR)的t(8；21)相关急性髓系白血病(AML)患者的AML1/ETO融合基因转录本。因此，定量测定CR期间融合转录本的数量可能比简单的定性评估更能预测治愈或复发。实时聚合酶链式反应是一种基于荧光的技术，与终点定量方法相比，它允许在聚合酶链式反应扩增的延伸阶段简单而快速地定量目标序列。对6例t(8；21)(q22；q22)AML患者在确诊及大剂量阿糖胞苷和蒽环类药物诱导治疗后不同时间段进行实时荧光定量RT-PCR检测。5名患者的诊断骨髓(BM)样本可用于分子分析。在诊断时，每个患者的AML1/ETO融合转录本的拷贝数都大于或等于10(3)个拷贝，并且在成功诱导化疗后，每个患者的拷贝数都减少了2到4个对数。这与白血病原始细胞的对数倍减少相当，后者被认为发生在通过诱导化疗成功将细胞减少为CR的患者中。第6例患者在成功的诱导缓解化疗后立即表现出相对较高的拷贝数，在早期CR期间继续增加，随后复发。与以前使用的定量RT-PCR方法相比，实时RT-PCR似乎提供了优势，因为它提供了目标序列的绝对定量，将定量的动态范围扩展到超过六个数量级，消除了PCR后处理，并减少了劳动力和残留污染。这些特征使其成为前瞻性评估AML1/ETO融合转录本定量在t(8；21)(q22；q22)AML患者中的预后价值的一个有吸引力的方法。
The AML1/ETO fusion transcript can be detected by reverse transcription polymerase chain reaction (RT-PCR) in patients with t(8;21)-associated acute myeloid leukemia (AML) in longterm complete remission (CR). Quantitation of the amount of the fusion transcript during CR may therefore be more predictive of cure or relapse than a simple qualitative assessment. Real Time PCR, a fluorometric-based technique, allows simple and rapid quantitation of a target sequence during the extension phase of PCR amplification, in contrast to end-point quantitative methods. Six patients with t(8;21)(q22;q22) AML, who achieved CR were studied by Real Time RT-PCR at different time intervals following diagnosis and high-dose cytarabine and anthracycline-based induction therapy. Five patients had a diagnostic bone marrow (BM) sample available for molecular analysis. Each patient showed greater than or equal to 10(3) copies of the AML1/ETO fusion transcript at diagnosis, and each showed a 2- to 4-log decrease in copy number following successful induction chemotherapy. This is comparable to the log-fold reduction in leukemic blasts that is thought to occur in patients successfully cytoreduced into CR by induction chemotherapy. The sixth patient showed a relatively high copy number immediately following successful remission induction chemotherapy, which continued to increase during early CR and was later followed by relapse. Real Time RT-PCR appears to offer advantages over previously used quantitative RT-PCR methods by providing absolute quantitation of the target sequence, expanding the dynamic range of quantitation to over six orders of magnitude, eliminating the post-PCR processing, and reducing labor and carryover contamination. These features make this an attractive method to prospectively evaluate the prognostic value of AML1/ETO fusion transcript quantitation in a larger patient population with t(8;21)(q22;q22) AML in CR.