Flow Cytometry Clinical Laboratory, Hospital Isarelita Albert Einstein, São Paulo, Brazil.
Cytometry B Clin Cytom. 2020 Nov;98(6):529-535. doi: 10.1002/cyto.b.21875. Epub 2020 Apr 6.
Minimal residual disease (MRD) in chronic lymphocytic leukemia (CLL) has prognostic and predictive significance. One of the approaches to detect MRD by flow cytometry (FC) is the use of dry antibody reagents such as DuraClone® RE CLB (Beckman Coulter-BC). The aim of this study was to evaluate the performance of the DuraClone® RE CLB in detecting MRD in CLL compared to liquid reagents.
DuraClone® RE CLB is composed by CD81FITC, ROR1PE, CD79bPC5.5, CD19PC7, CD5APC, CD43APCA750, CD20PB, and CD45KrO. For the liquid reagent assay, we used CD43FITC, ROR1PE, CD3ECD, CD5PC5.5, CD20PC7, CD79bAPC, CD19APC750, CD81 APCH7, and CD45KrO. The liquid and dry tubes were used to detect 20 MRD-positive CLL samples. The samples were analyzed using Radar Plots Kaluza Software (BC).
The statistical correlation between the liquid and dry reagents was acceptable (R = .9583) and no discrepancy was observed in MRD percentages. The average of the total number of acquired events in DuraClone® RE CLB was 758.583 (362.632-2.290.387), which allowed accurate sensitivity for the FC assay. The lowest MRD frequency detected by DuraClone® RE CLB was 0.01%, corresponding to a cluster with 106 events in a total of 737.030. The radar plots allowed the discrimination between normal B-cell population and CLL cells.
The DuraClone® RE CLB method allowed the accurate detection of MRD in clinical and interlaboratorial CLL samples, thereby supporting the use of this method to potentially increase productivity, reduce pipetting-associated errors and cost, and allow better standardization.
慢性淋巴细胞白血病(CLL)中的微小残留病(MRD)具有预后和预测意义。通过流式细胞术(FC)检测 MRD 的一种方法是使用干燥抗体试剂,如 DuraClone® RE CLB(贝克曼库尔特-BC)。本研究的目的是评估 DuraClone® RE CLB 在检测 CLL 中的 MRD 方面的性能,与液体试剂相比。
DuraClone® RE CLB 由 CD81FITC、ROR1PE、CD79bPC5.5、CD19PC7、CD5APC、CD43APCA750、CD20PB 和 CD45KrO 组成。对于液体试剂检测,我们使用 CD43FITC、ROR1PE、CD3ECD、CD5PC5.5、CD20PC7、CD79bAPC、CD19APC750、CD81 APCH7 和 CD45KrO。使用液体和干燥管检测 20 个 MRD 阳性的 CLL 样本。使用雷达图 Kaluza 软件(BC)对样本进行分析。
液体和干燥试剂之间的统计相关性是可以接受的(R =.9583),并且在 MRD 百分比方面没有观察到差异。在 DuraClone® RE CLB 中获得的总事件数的平均值为 758.583(362.632-2.290.387),这使得 FC 检测具有准确的灵敏度。DuraClone® RE CLB 检测到的最低 MRD 频率为 0.01%,对应于一个包含 106 个事件的簇,总共 737.030 个事件。雷达图允许区分正常 B 细胞群和 CLL 细胞。
DuraClone® RE CLB 方法允许准确检测临床和实验室间 CLL 样本中的 MRD,从而支持使用该方法来提高生产力、减少与移液相关的错误和成本,并更好地标准化。