Chelysheva E Iu, Turkina A G, Misiurin A V, Aksenova E V, Domracheva E V, Zakharova A V, Khoroshko N D
Ter Arkh. 2007;79(4):49-53.
To quantitatively determine minimal residual disease (MRD) by real-time polymerase chain reaction (PCR) in patients with a chronic phase (CP) of chronic myeloid leukemia (CML).
A molecular response was analyzed in 53 CML CP patients with incomplete and complete cptogenetic response (ICR and CCR) during imatinib therapy (median follow-up 36 months). BCR-ABL gene type p210 expression was quantitatively determined by real-time PCR under the TaqMan technology (an ICycler IQ device). The beta2 microglobulin (beta2M) gene was used as a reference gene. The results were expressed as the ratio: the number of BCR-ABL copies to that of beta2M x 10(5), as well as the difference of the common logarithm (lg) of the baseline expression level (BEL) and the result obtained: CEL lg-result lg.
The study revealed a correlation of the results of real-time PCR with those of cytogenetic analysis and showed it possible to study not only bone marrow, but also peripheral blood. Some negative real-time PCR results were checked using more sensitive PCR techniques. MRD was identified in most CML patients showing ICR and CCR during imatinib therapy. The reduction in BCR-ABL transcript levels by less than 2 lg (as compared to BEL) was associated with a cytogenetic recurrence and that by less than 3 lg was associated with a permanent high cytogenetic response. In patients with a cytogenetic recurrence, the median of BCR-ABL transcript levels was higher than that in patients with a permanent stable or unstable cytogenetic response. An elevation of BCR-ABL transcript levels over time antedated the development of a cytogenetic recurrence.
Quantitative monitoring by real-time PCR gives additional information on the dynamics of MRD in CML patients treated with glivec and permits improvement of study protocols for patients with CML at complete clinicohematological and cytogenetic remission.
通过实时聚合酶链反应（PCR）定量检测慢性髓性白血病（CML）慢性期（CP）患者的微小残留病（MRD）。
对53例接受伊马替尼治疗（中位随访36个月）且处于不完全和完全细胞遗传学反应（ICR和CCR）的CML CP患者进行分子反应分析。采用TaqMan技术（ICycler IQ仪器）通过实时PCR定量检测BCR-ABL基因p210型的表达。β2微球蛋白（β2M）基因用作参照基因。结果以BCR-ABL拷贝数与β2M×10(5)拷贝数之比表示，同时也表示为基线表达水平（BEL）与所得结果的常用对数（lg）之差：CEL lg - 结果lg。
该研究揭示了实时PCR结果与细胞遗传学分析结果之间的相关性，并表明不仅可以研究骨髓，还可以研究外周血。部分实时PCR阴性结果采用更敏感的PCR技术进行了验证。多数在伊马替尼治疗期间表现为ICR和CCR的CML患者检测到MRD。与BEL相比，BCR-ABL转录水平降低少于2 lg与细胞遗传学复发相关，降低少于3 lg与持久的高细胞遗传学反应相关。在细胞遗传学复发的患者中，BCR-ABL转录水平的中位数高于细胞遗传学反应持久稳定或不稳定的患者。随着时间推移BCR-ABL转录水平升高先于细胞遗传学复发的发生。
实时PCR定量监测为接受格列卫治疗的CML患者MRD的动态变化提供了额外信息，并有助于改进处于完全临床血液学和细胞遗传学缓解期的CML患者的研究方案。
