Objective: To compare the sensitivity of 8-color panels and next generation flow cytometry (NGF) for detecting minimal residual disease of multiple myeloma patients. Methods: 8-color-membrane antigens (8C-Mem) panel was built including CD45, CD38, CD138, CD19, CD56, CD81, CD27 and CD117 to identify the plasma cells, while 8-color-cytoplasmic antigens (8C-Cyto) panel was built including CD45, CD38, CD138, CD19, CD56, CD81, cKappa (cK) and cLambda (cλ) , and 8-color-two-tubes (8C-2tubes) panel were built including 8C-Mem and 8C-Cyto panels, the data of three groups was analyzed by Diva software. NGF uses Infinicyt software to fuse 8C-2tubes data to further analyze the expression of plasma antigens. Bone marrow aspiration obtained from 20 controls and 76 multiple myeloma patients who achieved complete remission were measured and analyzed. Results: Positive MRD samples were discriminated in 88.2% of the specimen evaluated through either abnormal plasma cells (aPCs) or clonal plasma cells (cPCs) by NGF antigens panel, Among of them, consistency was 94.7%. The median percentage of cPCs was 0.3530%, The lowest sensitivity of NGF was 0.0003%. In 8-color panels, the positive MRD rates of 8C-Mem, 8C-Cyto and 8C-2tubes panels were 84.2%, 85.5% and 86.8%, respectively, which lower than that of NGF (P<0.001) . The positive MRD rate of 8C-Mem and 8C-Cyto panels were lower than that of 8C-2tubes panel (P<0.001) , and the positive MRD rate of 8C-Mem panel was lower than that of 8C-Cyto panel (P<0.001) . Sensitivity and specificity of NGF was higher than that of 8-color panels. 8C-2tubes panel has the best sensitivity, accuracy, negative predicted value, positive predicted value and specificity than other 8-color panels. However, huge data and low efficiency for analysis is the disadvantage. 8C-Cyto panel was the second choice, and 8C-Mem panel was the last. Conclusions: Membrane and cytoplasmic light chain is a better method for multiple myeloma-MRD detection and NGF panel is an ideal approach. 8C-Cyto panel is recommended in 8-MFC groups.
目的： 比较8色组合和二代流式细胞术（NGF）检测多发性骨髓瘤（MM）患者治疗后微小残留病（MRD）的敏感性。 方法： 分别建立一组检测胞膜抗原CD45、CD38、CD138、CD19、CD56、CD117、CD81和CD27的8C-Mem组，及一组检测胞膜及胞质抗原CD45、CD38、CD138、CD19、CD56、CD27、cκ和cλ的8C-Cyto组，将8C-Mem组和8C-Cyto组合成的8C-2tubes组，3组采用Diva软件分析浆细胞胞膜抗原表达与胞质轻链表达，统称8色组合；NGF是采用Infinicyt软件将8C-2tubes组两管数据融合，进一步分析浆细胞各抗原的表达。以20名正常对照和76例治疗后达完全缓解的MM患者骨髓样本进行检测与比较分析。 结果： NGF组合识别异常浆细胞（aPC）和克隆性浆细胞（cPC）的阳性率均为88.2%（67/76），一致率为94.7%（72/76），cPC的中位值为0.3530%，均为胞质轻链限制性表达，NGF检测的敏感度为0.0003%。8色组合中，8C-Mem组、8C-Cyto组和8C-2tubes组MRD阳性率分别为84.2%、85.5%和86.8%，均低于NGF的88.2%（P<0.001）；8C-Cyto组和8C-Mem组MRD阳性率低于8C-2tubes组（P<0.001），8C-Mem组MRD阳性率低于8C-Cyto组（P<0.001）。NGF敏感性和特异性优于8色组合，8色组合中8C-2tubes组敏感性和阴性预测值、特异性和阳性预测值及准确性最高，缺点是获取数据太大，采用Diva软件分析耗时长，效率低；其次是8C-Cyto组，8C-Mem最低。 结论： 反应浆细胞克隆性的胞质轻链较胞膜抗原更特异，胞膜抗原异常表达不能完全代表cPC，建议胞膜抗原和胞质轻链同时检测，有条件者优先选择NGF，8色组合中建议选择8C-Cyto组。.
Keywords: Minimal Residual disease; Multiple myeloma; Next generation flow cytometry.