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利用含冻干抗体试剂的八色管检测 B 细胞急性淋巴细胞白血病的微小残留病。

Detection of Minimal Residual Disease in B Cell Acute Lymphoblastic Leukemia Using an Eight-Color Tube with Dried Antibody Reagents.

机构信息

Assistance Publique Hôpitaux de Marseille, Laboratoire d'Hématologie, Hôpital de la Timone, Marseille, France.

CNRS, INSERM, CIML, Aix Marseille University, Marseille, France.

出版信息

Cytometry B Clin Cytom. 2019 Mar;96(2):158-163. doi: 10.1002/cyto.b.21766. Epub 2019 Jan 30.

Abstract

BACKGROUND

Flow cytometry is a powerful tool for the detection of minimal residual disease (MRD) of B cell precursor acute lymphoblastic leukemia (BCP-ALL) patients. However, the staining process and the choice of antibodies rely on laboratory expertise and may be source of variability or technical errors. Recently, Beckman Coulter commercialized a ready to use tube with dried format reagents for BCP-ALL MRD detection. The aim of this study is to evaluate the applicability of this tube and to compare it to a conventional (liquid format reagents) method.

METHODS

Thirty-one samples from B ALL patients were analyzed: 19 bone marrow (BM) aspirations, 10 peripheral blood (PB) samples and 2 cerebrospinal fluids at different stages of the follow-up. In addition, we tested 5 bone marrow samples mixed into non-pathological (control) bone marrow. The dried format tube included seven antibodies: CD45Kro, CD58FITC, CD34ECD, CD10PC5.5, CD19PC7, CD38AA700, CD20AA750, with possibility of additional antibodies for blast markers identified at diagnosis. For comparison, a liquid format tube was prepared, and considered as the reference.

RESULTS

This tube was validated for daily routine laboratory, with satisfying qualitative (MRD + or MRD-) and quantitative (MRD percentages) correlation with the reference tube.

CONCLUSION

With this single dried format tube, we showed interesting results for BCP-ALL MRD detection in the aim of standardization and reliable interlaboratory results. It allows accurate MRD detection including low levels (10-4), and offers possibility to increase performance (supplementary antibody) within a preestablished effective antibody panel for BCP-ALL MRD. © 2018 International Clinical Cytometry Society.

摘要

背景

流式细胞术是检测 B 细胞前体急性淋巴细胞白血病(BCP-ALL)患者微小残留病(MRD)的有力工具。然而,染色过程和抗体的选择依赖于实验室的专业知识,可能是变异或技术误差的来源。最近,贝克曼库尔特公司推出了一种用于 BCP-ALL MRD 检测的即用型干试剂管。本研究旨在评估该管的适用性,并将其与传统(液体试剂)方法进行比较。

方法

分析了 31 例 B-ALL 患者的样本:19 例骨髓(BM)穿刺、10 例外周血(PB)样本和 2 例不同随访阶段的脑脊液。此外,我们还测试了 5 例混入非病理(对照)骨髓的骨髓样本。干试剂管包括七种抗体:CD45Kro、CD58FITC、CD34ECD、CD10PC5.5、CD19PC7、CD38AA700、CD20AA750,并可根据诊断时确定的 blast 标志物添加额外的抗体。为了比较,我们制备了一个液体试剂管,并将其作为参考。

结果

该管经过验证可用于日常实验室,与参考管具有满意的定性(MRD+或 MRD-)和定量(MRD 百分比)相关性。

结论

使用这种单一的干试剂管,我们在标准化和可靠的实验室间结果方面显示了 BCP-ALL MRD 检测的有趣结果。它允许进行准确的 MRD 检测,包括低水平(10-4),并提供了在用于 BCP-ALL MRD 的预定义有效抗体组内增加性能(补充抗体)的可能性。 © 2018 国际临床细胞化学学会。

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