Han Lan-xiu, Lin Jiang, Qian Jun, Wang Ya-li, Qian Zhen, Yang Xiao-fei, Sheng Xiao-jing, Yao Dong-ming
Department of Hematology, Affiliated People's Hospital of Jiangsu University, Zhenjiang 212002, China.
Zhonghua Xue Ye Xue Za Zhi. 2008 Nov;29(11):753-6.
To establish the real time quantitative PCR (RQ-PCR) assay according to 'Europe Against Cancer' (EAC) program and analyze the results of detection and quantification of different PML-RAR alpha transcript isoforms in patients with acute promyelocytic leukemia (APL).
Three RQ-PCR systems were performed to detect the most frequent PML-RAR alpha transcripts (L-form, S-form and V-form) in 30 APL patients and the RQ-PCR end-point products were identified by electrophoresis.
S-form RQ-PCR system could amplify positive signals of three isoforms in all of 30 cases, and V-form RQ-PCR system could do so in both L-form and V-form positive cases, however, L-form RQ-PCR system could only do so in L-form-positive cases. Electrophoresis and sequencing of end-point products amplified by S-form RQ-PCR system revealed three bands in each of L-form (621 bp, 477 bp and 218 bp) and V-form (567 bp, 423 bp and 218 bp) positive patients samples.
RQ-PCR, sensitive and reliable, can be used for monitoring the minimal residual disease in APL patients, however, its results should be interpreted carefully if it is used for detection of PML-RAR alpha fusion transcripts prospectively.
按照“欧洲抗癌计划”（EAC）建立实时定量聚合酶链反应（RQ-PCR）检测方法，并分析急性早幼粒细胞白血病（APL）患者不同PML-RARα转录本异构体的检测和定量结果。
采用三种RQ-PCR系统检测30例APL患者中最常见的PML-RARα转录本（L型、S型和V型），并通过电泳鉴定RQ-PCR终点产物。
S型RQ-PCR系统在30例患者中均能扩增出三种异构体的阳性信号，V型RQ-PCR系统在L型和V型阳性病例中均能扩增出阳性信号，而L型RQ-PCR系统仅在L型阳性病例中能扩增出阳性信号。S型RQ-PCR系统扩增的终点产物电泳和测序显示，L型（621 bp、477 bp和218 bp）和V型（567 bp、423 bp和218 bp）阳性患者样本中各出现三条条带。
RQ-PCR敏感可靠，可用于监测APL患者的微小残留病，但前瞻性检测PML-RARα融合转录本时，结果应谨慎解读。
