Moumen A, Dehbi H, Kottwitz D, El Amrani M, Bouchoutrouch N, El Hadi H, Quessar A, Benchekroun S, Nadifi S, Sefrioui H
Moroccan Foundation for Advanced Science, Innovation and Research Rabat, Morocco.
Laboratory of Human Genetics, Faculty of Medicine, Casablanca, Morocco.
Genet Mol Res. 2015 Feb 6;14(1):1044-55. doi: 10.4238/2015.February.6.8.
Chronic myeloid leukemia (CML) is characterized by BCR-ABL translocation and an increased number and migration of immature myeloid cells into the peripheral blood. The detection limit of the BCR-ABL transcript, particularly after treatment, is controversial. In the present study, we used quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR) to monitor BCR-ABL expression in Moroccan CML patients undergoing imatinib treatment, and compared the results with those of conventional PCR and fluorescence in situ hybridization (FISH). The aim of this study was to establish a new molecular tool for in vitro diagnosis of CML. In a retrospective comparative analysis, 20 CML Moroccan patients who had received imatinib treatment (N = 20) were analyzed by real-time PCR, conventional RT-PCR, and FISH. Half of the samples analyzed (N = 10) were positive for BCR-ABL gene expression, while the other half (N = 10) were negative according to conventional PCR. Interestingly, 5 of the 10 samples shown to be negative by conventional PCR showed positive expression of the BCR-ABL gene according to RT-qPCR. The RT-qPCR results were confirmed by FISH, which revealed a high concordance (100%) rate. We found that real-time RT-qPCR is more reliable and should be used in Moroccan biomedical analysis laboratories to monitor CML progression, particularly for minimal residual disease, following imatinib treatment.
慢性髓性白血病(CML)的特征是BCR-ABL易位以及未成熟髓细胞数量增加并迁移至外周血。BCR-ABL转录本的检测限,尤其是在治疗后,存在争议。在本研究中,我们使用定量实时逆转录-聚合酶链反应(RT-qPCR)来监测接受伊马替尼治疗的摩洛哥CML患者的BCR-ABL表达,并将结果与传统PCR和荧光原位杂交(FISH)的结果进行比较。本研究的目的是建立一种用于CML体外诊断的新分子工具。在一项回顾性比较分析中,对20例接受伊马替尼治疗的摩洛哥CML患者(N = 20)进行了实时PCR、传统RT-PCR和FISH分析。根据传统PCR,所分析样本的一半(N = 10)BCR-ABL基因表达呈阳性,而另一半(N = 10)呈阴性。有趣的是,在传统PCR显示为阴性的10个样本中,有5个根据RT-qPCR显示BCR-ABL基因呈阳性表达。RT-qPCR结果通过FISH得到证实,FISH显示出高一致性(100%)率。我们发现实时RT-qPCR更可靠,应在摩洛哥生物医学分析实验室中用于监测CML进展,特别是在伊马替尼治疗后用于检测微小残留病。