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[四色和八色荧光抗体组合检测多发性骨髓瘤患者微小残留病的比较与分析]

[Comparison and analysis of minimal residual disease in multiple myeloma patients detected by 4-color and 8-color fluorescence antibody panels].

作者信息

Wang Y Z, Lu J, Hao L, Chang Y, He L L, Huang X J, Liu Y R

机构信息

Peking University Institute of Hematology, People's Hospital of Peking University, Beijing Key Laboratory of Hematopoietic Stem Cell Transplantation, Beijing 100044, China.

出版信息

Zhonghua Xue Ye Xue Za Zhi. 2017 Apr 14;38(4):272-278. doi: 10.3760/cma.j.issn.0253-2727.2017.04.003.

Abstract

To explore and compare 4-color and 8-color fluorescence antibody panels for detecting minimal residual disease of multiple myeloma patients after therapy. One 8-color antibody panel was built including CD38 and CD138 for the identification of plasma cells (PCs) , membrane antigen CD45, CD19, CD56 and CD117, cytoplasmic Kappa (cκ) and Lambda (cλ) light chain antigen. Six tubes of 4-color panels were built, among them, membrane antigen CD45/CD19, CD56/CD117, CD19/CD56 and light chains were analyzed combined by CD38/CD138 for PCs gate in the tubes M1-3 and tube C-(κ/λ), respectively; CD19 or CD45 and cκ/cλ light chains were detected in the tube MC1-(C)D38 for CD38/SSC identified PCs gate and tube MC2-(CD138) for CD138/SSC identified PCs gate separately. Twenty normal volunteer bone marrows and seventy-three specimens from multiple myeloma patients after complete remission were measured and analyzed. MRD positive samples were discriminated in 82.19% of the specimen evaluated through either abnormal plasma cells (aPCs) or clonal plasma cells (cPCs) by 8-color antibody panel. Among of them, consistency was 89.04%. The median percentage of cPCs was 0.105%. The lowest sensitivity of experiment was 0.004%. Percentage of PCs identified by CD38/ SSC gate was higher than that by CD38/CD138 (<0.001) and CD138/SSC gate (=0.001) apparently. The lowest MRD positive rate was found in tube C (65.75%) , which lower than 8-color panel obviously (=0.024) . The percentages of aPCs measured by tube M2-(56/117) were significantly lower than other tubes (=0.014) . MRD positive rate determined by cPCs was higher than that by aPCs both in the tube MC1-(C)D38 and tube MC2-(CD138), whose concordance rate were 68.49% and 79.45%, respectively. Compared with 8-color panel, tube MC2-(CD138) the best choice among six tubes of 4-color panels, which has the best sensitivity, accuracy and negative predicted value, higher positive predicted value and specificity. Tube M1-(45/19) was the second choice. CD38/CD138 are the best markers for gating PCs. Membrane antigen and cκ/cλ detected simultaneously is a better method for MM MRD detection and 8-color antibody panel is an ideal approach. Two tubes of 4-color antibody combination, M1-(45/19) and MC2-(CD138) are recommended in the 4-color panels.

摘要

探索并比较4色和8色荧光抗体组合用于检测多发性骨髓瘤患者治疗后的微小残留病。构建了一个8色抗体组合,包括用于识别浆细胞(PCs)的CD38和CD138、膜抗原CD45、CD19、CD56和CD117、细胞质κ(cκ)和λ(cλ)轻链抗原。构建了6管4色组合,其中,在管M1 - 3和管C -(κ/λ)中,分别通过CD38/CD138对PCs门控,联合分析膜抗原CD45/CD19、CD56/CD117、CD19/CD56和轻链;在管MC1 -(C)D38中针对CD38/SSC识别的PCs门控检测CD19或CD45以及cκ/cλ轻链,在管MC2 -(CD138)中针对CD138/SSC识别的PCs门控分别检测。对20份正常志愿者骨髓和73份多发性骨髓瘤患者完全缓解后的标本进行检测和分析。通过8色抗体组合,在评估的标本中,82.19%的标本通过异常浆细胞(aPCs)或克隆浆细胞(cPCs)鉴别出微小残留病阳性样本。其中,一致性为89.04%。cPCs的中位百分比为0.105%。实验的最低灵敏度为0.004%。CD38/SSC门控识别的PCs百分比明显高于CD38/CD138(<0.001)和CD138/SSC门控(=0.001)。在管C中发现最低的微小残留病阳性率(65.75%),明显低于8色组合(=0.024)。管M2 -(56/117)检测的aPCs百分比显著低于其他管(=0.014)。在管MC1 -(C)D38和管MC2 -(CD138)中,通过cPCs确定的微小残留病阳性率均高于通过aPCs确定的,其符合率分别为68.49%和79.45%。与8色组合相比,管MC2 -(CD138)是4色组合6管中最佳选择,具有最佳的灵敏度、准确性和阴性预测值,较高的阳性预测值和特异性。管M1 -(45/19)是第二选择。CD38/CD138是门控PCs的最佳标志物。同时检测膜抗原和cκ/cλ是多发性骨髓瘤微小残留病检测的较好方法,8色抗体组合是理想的方法。在4色组合中推荐两管4色抗体组合,即M1 -(45/19)和MC2 -(CD138)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1bab/7342730/db5c5668042a/cjh-38-04-272-g001.jpg

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