Tang Bao-lin, Cheng Min, Liu Hui-lan, Geng Liang-quan, Wu Zhen-tian, Sun Zi-min
Department of Hematology, Anhui Provincial Hospital, being Affiliated to Anhui Medical University, Hefei 230001, China.
Zhonghua Nei Ke Za Zhi. 2007 Mar;46(3):224-8.
To monitor the gene expression of BCR/ABL in chronic myeloid leukemia patients after allogeneic hematopoietic stem cells transplantation with real-time RT-PCR for evaluating therapeutic efficacy and guiding further treatment.
Taqman real-time RT-PCR technique was performed to monitor peripheral blood BCR/ABL transcript levels in 10 patients at the time of diagnosis and serially at intervals in 25 patients after allogeneic hematopoietic stem cells transplantation. beta-actin mRNA was used as an endogenous reference.
The sensitivity of real-time RT-PCR was 10 copies/microl. In 15 patients receiving transplantation, the median of N(BCR/ABL) (%) before transplantation, 1 month after transplantation and 2 months after transplantation was respectively: 6.57 (0.14 - 38.83), 0.10 (0 - 1.71) and 0 (0 - 0.52). 3 months later N(BCR/ABL) (%) all turned to 0. BCR/ABL transcripts after transplantation were detected to decrease gradually. N(BCR/ABL) (%) 1 month after transplantation and 2 months after transplantation was both lower than that before transplantation (chi(2) both = 13.07, P < 0.01). N(BCR/ABL) (%) at 2 months after transplantation was lower than that at 1 month after transplantation (chi(2) = 8.10, P < 0.01). In 10 other patients, N(BCR/ABL) (%) was 0 in consecutive tests from 3 to 43 months after transplantation.
Real-time RT-PCR is sensitive, reliable and reproducible for monitoring CML relapse after transplantation. It is of help in detecting minimal residual disease, predicting the prognosis of the disease and providing practical indications for further treatment.
采用实时逆转录聚合酶链反应(RT-PCR)监测异基因造血干细胞移植后慢性髓性白血病患者BCR/ABL的基因表达,以评估治疗效果并指导进一步治疗。
运用Taqman实时RT-PCR技术,在诊断时对10例患者的外周血BCR/ABL转录水平进行监测,并在25例异基因造血干细胞移植后的患者中定期进行连续监测。β-肌动蛋白mRNA用作内参。
实时RT-PCR的灵敏度为10拷贝/微升。15例接受移植的患者中,移植前、移植后1个月和移植后2个月的N(BCR/ABL)(%)中位数分别为:6.57(0.14 - 38.83)、0.10(0 - 1.71)和0(0 - 0.52)。3个月后N(BCR/ABL)(%)均转为0。移植后检测到BCR/ABL转录本逐渐减少。移植后1个月和移植后2个月的N(BCR/ABL)(%)均低于移植前(χ²均 = 13.07,P < 0.01)。移植后2个月的N(BCR/ABL)(%)低于移植后1个月(χ² = 8.10,P < 0.01)。另外10例患者在移植后3至43个月的连续检测中N(BCR/ABL)(%)均为0。
实时RT-PCR对于监测移植后慢性髓性白血病复发敏感、可靠且可重复。有助于检测微小残留病,预测疾病预后并为进一步治疗提供实际指导。