Evaluation of candidate control genes for diagnosis and residual disease detection in leukemic patients using 'real-time' quantitative reverse-transcriptase polymerase chain reaction (RQ-PCR) - a Europe against cancer program
Evaluation of candidate control genes for diagnosis and residual disease detection in leukemic patients using 'real-time' quantitative reverse-transcriptase polymerase chain reaction (RQ-PCR) - a Europe against cancer program
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DOI：
10.1038/sj.leu.2403136
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发表时间：
2003-12-01
期刊：
影响因子：
11.4
通讯作者：
Gabert, J
中科院分区：
文献类型：
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作者：
Beillard, E;Pallisgaard, N;Gabert, J
实时定量RT-PCR（RQ-PCR）是一种通过扩增融合基因（FG）转录本来监测白血病患者微小残留病（MRD）的敏感工具。为了校正RNA质量和数量的变化并计算每次测量的灵敏度，应与FG转录物平行扩增对照基因（CG）转录物。为了确定合适的CG，欧洲抗癌（EAC）计划中的一个研究小组最初使用标准化的RQ-PCR方案集中在14个潜在的CG上。基于不存在假基因以及CG表达的水平和稳定性，最终选择了三个基因：Abelson（ABL）、β-2-微球蛋白（B2 M）和β-葡萄糖醛酸酶（GUS）。对同一天处理的正常（n = 126）和诊断性白血病（n = 184）样本进行的多中心前瞻性研究已经建立了CG表达的参考值。对诊断时获得的超过250个急性和慢性白血病样本进行的多中心回顾性研究证实，这三种CG在不同类型的样本中具有稳定的表达。然而，只有ABL基因转录表达没有显着差异正常和白血病样本在诊断。因此，我们建议使用ABL基因作为CG的RQ-PCR为基础的诊断和MRD检测白血病患者。总之，这些数据不仅适合于融合基因转录本的定量，而且适合于异常表达基因的定量。
Real-time quantitative RT-PCR (RQ-PCR) is a sensitive tool to monitor minimal residual disease (MRD) in leukemic patients through the amplification of a fusion gene (FG) transcript. In order to correct variations in RNA quality and quantity and to calculate the sensitivity of each measurement, a control gene (CG) transcript should be amplified in parallel to the FG transcript. To identify suitable CGs, a study group within the Europe Against Cancer (EAC) program initially focused on 14 potential CGs using a standardized RQ-PCR protocol. Based on the absence of pseudogenes and the level and stability of the CG expression, three genes were finally selected: Abelson (ABL), beta-2-microglobulin (B2M), and beta-glucuronidase (GUS). A multicenter prospective study on normal (n = 126) and diagnostic leukemic ( n = 184) samples processed the same day has established reference values for the CG expression. A multicenter retrospective study on over 250 acute and chronic leukemia samples obtained at diagnosis and with an identified FG transcript confirmed that the three CGs had a stable expression in the different types of samples. However, only ABL gene transcript expression did not differ significantly between normal and leukemic samples at diagnosis. We therefore propose to use the ABL gene as CG for RQ-PCR-based diagnosis and MRD detection in leukemic patients. Overall, these data are not only eligible for quantification of fusion gene transcripts, but also for the quantification of aberrantly expressed genes.