Department of Immunology, Erasmus MC, University Medical Center, Rotterdam, The Netherlands.
Leukemia. 2013 Mar;27(3):635-41. doi: 10.1038/leu.2012.231. Epub 2012 Aug 16.
Most current treatment protocols for acute lymphoblastic leukemia (ALL) include minimal residual disease (MRD) diagnostics, generally based on PCR analysis of rearranged antigen receptor genes. Although flow cytometry (FCM) can be used for MRD detection as well, discordant FCM and PCR results are obtained in 5-20% of samples. We evaluated whether 6-color FCM, including additional markers and new marker combinations, improved the results. Bone marrow samples were obtained from 363 ALL patients at day 15, 33 and 78 and MRD was analyzed using 6-color (218 patients) or 4-color (145 patients) FCM in parallel to routine PCR-based MRD diagnostics. Compared with 4-color FCM, 6-color FCM significantly improved the concordance with PCR-based MRD data (88% versus 96%); particularly the specificity of the MRD analysis improved. However, PCR remained more sensitive at levels <0.01%. MRD-based risk groups were similar between 6-color FCM and PCR in 68% of patients, most discrepancies being medium risk by PCR and standard risk by FCM. Alternative interpretation of the PCR data, aimed at prevention of false-positive MRD results, changed the risk group to standard risk in half (52%) of these discordant cases. In conclusion, 6-color FCM significantly improves MRD analysis in ALL but remains less sensitive than PCR-based MRD-diagnostics.
目前大多数急性淋巴细胞白血病 (ALL) 的治疗方案都包括微小残留病 (MRD) 诊断，通常基于抗原受体基因重排的 PCR 分析。虽然流式细胞术 (FCM) 也可用于 MRD 检测，但在 5-20%的样本中会得到与 FCM 和 PCR 结果不一致的结果。我们评估了 6 色 FCM 是否能通过增加额外的标记物和新的标记物组合来改善结果。从 363 例 ALL 患者的第 15、33 和 78 天的骨髓样本中获得了骨髓样本，并通过 6 色（218 例患者）或 4 色（145 例患者）FCM 与基于常规 PCR 的 MRD 诊断平行进行 MRD 分析。与 4 色 FCM 相比，6 色 FCM 显著提高了与基于 PCR 的 MRD 数据的一致性（88% 对 96%）；特别是 MRD 分析的特异性得到了改善。然而，在 <0.01%的水平上，PCR 仍然更敏感。在 68%的患者中，6 色 FCM 和 PCR 之间的基于 MRD 的风险组相似，最主要的差异是 PCR 为中危风险，而 FCM 为标准风险。针对 PCR 数据中假阳性 MRD 结果的解读，将风险组更改为标准风险，在这些不一致的病例中有一半（52%）。总之，6 色 FCM 显著改善了 ALL 中的 MRD 分析，但仍不如基于 PCR 的 MRD 诊断敏感。
