Schoch C, Schnittger S, Bursch S, Gerstner D, Hochhaus A, Berger U, Hehlmann R, Hiddemann W, Haferlach T
Department of Internal Medicine III, University Hospital Grosshadern, Ludwig-Maximilians-University, Munich, Germany.
Leukemia. 2002 Jan;16(1):53-9. doi: 10.1038/sj.leu.2402329.
For the diagnosis of CML and for monitoring of treatment response the detection of the t(9;22)(q34;q11) or the BCR-ABL rearrangement is necessary. Chromosome banding analysis (CA) is still the gold standard but other techniques like Southern blot, fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR) are available. We analyzed 350 CML patients at different stages of disease in parallel with CA, interphase-FISH (IP-FISH), hypermetaphase-FISH (HM-FISH) and RT-PCR. In 20 cases with no Ph(+) metaphases in CA, HM-FISH detected 0.2 to 10% BCR-ABL(+)metaphases. After IP-FISH 107 samples were judged as negative. However, in 17 of these samples HM-FISH detected BCR-ABL(+) metaphases (0.3-11%), and in eight cases CA detected Ph(+) metaphases (2.5-25%). A comparison of IP-FISH performed on uncultivated cells vs cells cultivated for 48 h in 70 cases revealed a higher proportion of BCR-ABL+ cells in the cultivated samples. If nested PCR was negative, all other methods were negative in all cases too. In addition, 94 cases were evaluated using real-time PCR (LightCycler technology). The BCR-ABL/cABL ratio measured showed a high correlation with all other methods. Interestingly, a wide range in the BCR-ABL/ABL ratio was observed especially in patients who showed 100% Ph-positive metaphases in CA. In conclusion, CA, IP-FISH, HM-FISH and real-time PCR give reliable results but differences due to measurement of different target structures have to be kept in mind when using these data for definition of remission status.
为诊断慢性粒细胞白血病（CML）并监测治疗反应，检测t(9;22)(q34;q11)或BCR-ABL重排是必要的。染色体显带分析（CA）仍是金标准，但也有其他技术可用，如Southern印迹法、荧光原位杂交（FISH）和聚合酶链反应（PCR）。我们对350例处于不同疾病阶段的CML患者同时采用CA、间期FISH（IP-FISH）、超中期FISH（HM-FISH）和逆转录PCR（RT-PCR）进行分析。在CA检测未发现Ph(+)中期的20例病例中，HM-FISH检测到0.2%至10%的BCR-ABL(+)中期。IP-FISH检测后，107份样本被判定为阴性。然而，在这些样本中的17份中，HM-FISH检测到BCR-ABL(+)中期（0.3% - 11%），在8例病例中，CA检测到Ph(+)中期（2.5% - 25%）。对70例未经培养的细胞与培养48小时的细胞进行IP-FISH比较，结果显示培养样本中BCR-ABL+细胞的比例更高。如果巢式PCR为阴性，所有其他方法在所有病例中也均为阴性。此外，采用实时PCR（LightCycler技术）对94例病例进行了评估。所测的BCR-ABL/cABL比值与所有其他方法显示出高度相关性。有趣的是，尤其是在CA检测显示100% Ph阳性中期的患者中，观察到BCR-ABL/ABL比值范围很广。总之，CA、IP-FISH、HM-FISH和实时PCR均能给出可靠结果，但在使用这些数据定义缓解状态时，必须牢记由于测量不同靶结构而导致的差异。
