4色与8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：王亚哲[1] 路瑾[1] 郝乐[1] 常艳[1] 贺玲玲[1] 黄晓军[1] 刘艳荣[1] 

机构地区：[1]北京大学人民医院、北京大学血液病研究所,造血干细胞移植治疗血液病北京市重点实验室,100044

出　　处：《中华血液学杂志》2017年第4期272-278,共7页Chinese Journal of Hematology

摘　　要：目的探讨与比较4色和8色荧光抗体组合检测多发性骨髓瘤（MM）患者治疗后微小残留病（MRD）的灵敏度。 方法建立一组以CD38/CD138对浆细胞进行设门，同时检测胞膜抗原CD45、CD19、CD56、CD117及胞质轻链cKappa、cLambda的8色抗体组合；6组4色抗体组合中，4组以CD38/CD138设门，分别分析CD45/CD19、CD56/CD117、CD19/CD56胞膜抗原（M1-3）和胞质轻链（C组）表达；MC1-2组分别以CD38/SSC和CD138/SSC设门，同时检测CD19或CD45胞膜抗原和胞质轻链。采用以上组合对20名健康志愿者和73例完全缓解后MM患者标本进行检测并进行比较分析。 结果8色组合识别异常浆细胞（aPCs）和克隆性浆细胞（cPCs）的阳性率均为82.19%，一致率为89.04%（65/73）；cPCs中位比例为0.105%，均显示明显的单克隆轻链；敏感度为0.004%。CD38/SSC设门的浆细胞比例明显高于CD38/CD138（P〈0.001）和CD138/SSC（P=0.001）设门。4色组合中C-胞质组MRD阳性率最低，仅为65.75%，明显低于8色组合的82.19%（P=0.024）；M2-56/117组检测的aPCs比例最低，为0.095%，与其他组相比差异有统计学意义（P=0.014）；MC1-CD38和MC2-CD138组，以cPCs作为判断标准时MRD阳性率均高于以aPCs为判断标准（P〈0.05），结果一致率分别为68.49%（50/73）和79.45%（58/73）。与8色组合相比，4色组合中MC2-CD138组敏感性、准确性和阴性预测值最高，阳性预测值和特异性也较高；其次是M1-45/19组。 结论CD38/CD138设门更准确，反映克隆性的胞质轻链检测较胞膜抗原更敏感，胞膜抗原异常表达并不完全代表为cPCs，建议胞膜抗原和胞质轻链同时检测。有条件者建议选用8色组合，4色组合中建议同时采用M1-45/19和MC2-CD138组合。Objective To explore and compare 4-color and 8-color fluorescence antibody panels for detecting minimal residual disease of multiple myeloma patients after therapy. Methods One 8-color antibody panel was built including CD38 and CD138 for the identification of plasma cells （PCs）, membrane antigen CD45, CD19, CD56 and CDll7, cytoplasmic Kappa（ck） and Lambda（c） 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-m, respectively; CD19 or CD45 and c/c light chains were detected in the tube MCl-cD38 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. Results 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 CD3 8/SSC gate was higher than that by CD38/CD138 （P〈0.001） and CD138/SSC gate （P=0.001） apparently. The lowest MRD positive rate was found in tube C （65.75%）, which lower than 8-color panel obviously （P=0.024）. The percentages of aPCs measured by tube M2-56/7 were significantly lower than other tubes （P=0.014）. MRD positive rate determined by cPCs was higher than that by aPCs both in the tube MCl-cD38 and tube MC2-cD38, whose concordance rate were 68.49% and 79.45%, respectively. Compared with 8-color panel, tube MC2-cD,38 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 M 1-45/19 was the second choice. Concl

关 键 词：多发性骨髓瘤 微小残留病 

分 类 号：R733.3[医药卫生—肿瘤]