Comparative analysis of minimal residual disease detection using four-color flow cytometry, consensus IgH-PCR, and quantitative IgH PCR in CLL after allogeneic and autologous stem cell transplantation
Comparative analysis of minimal residual disease detection using four-color flow cytometry, consensus IgH-PCR, and quantitative IgH PCR in CLL after allogeneic and autologous stem cell transplantation
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DOI：
10.1038/sj.leu.2403478
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发表时间：
2004-10-01
期刊：
影响因子：
11.4
通讯作者：
Kneba, M
中科院分区：
文献类型：
--
作者：
Böttcher, S;Ritgen, M;Kneba, M
临床上最适合检测慢性淋巴细胞白血病微小残留病（MRD）的方法仍有争议。我们前瞻性地比较了74例CLL患者干细胞移植（SCT）后158份血液样本的MRD评估，采用四色流式细胞术（MRD flow）与共识IgH-PCR和阿索IgH实时PCR（阿索IgH RQ-PCR）平行。在具有多克隆共有IgH-PCR模式的106个样品中的25个（23.6%）中，MRD流仍然检测到CLL细胞，证明流式细胞术的灵敏度高于使用共有IgH引物的PCR基因扫描。在92例样本中，14例（15.2%）平行分析的MRD流和阿索IgH RQ-PCR检测结果为阴性，而我们的流式细胞术检测结果为阳性，因此证明了使用阿索引物的RQ-PCR具有上级灵敏度。两种方法测量的定量MRD水平相关性良好（r = 0.93）。流式MRD检测和阿索IgH RQ-PCR同样适用于监测同种异体SCT后的MRD动力学，但PCR方法更早地检测到自体SCT后即将复发的情况。对影响流式细胞术检测MRD的灵敏度和特异性的因素进行分析，可以进一步改进该技术。
The clinically most suitable method for minimal residual disease (MRD) detection in chronic lymphocytic leukemia is still controversial. We prospectively compared MRD assessment in 158 blood samples of 74 patients with CLL after stem cell transplantation (SCT) using four-color flow cytometry ( MRD flow) in parallel with consensus IgH-PCR and ASO IgH real-time PCR ( ASO IgH RQ-PCR). In 25 out of 106 samples (23.6%) with a polyclonal consensus IgH-PCR pattern, MRD flow still detected CLL cells, proving higher sensitivity of flow cytometry over PCR-genescanning with consensus IgH-primers. Of 92 samples, 14 (15.2%) analyzed in parallel by MRD flow and by ASO IgH RQ-PCR were negative by our flow cytometric assay but positive by PCR, thus demonstrating superior sensitivity of RQ-PCR with ASO primers. Quantitative MRD levels measured by both methods correlated well ( r = 0.93). MRD detection by flow and ASO IgH RQ-PCR were equally suitable to monitor MRD kinetics after allogeneic SCT, but the PCR method detected impending relapses after autologous SCT earlier. An analysis of factors that influence sensitivity and specificity of flow cytometry for MRD detection allowed to devise further improvements of this technique.