基于CD157的四色法流式细胞术单核细胞和中性粒细胞PNH克隆检测方法的建立及评价

Establishment and evaluation of CD157-based 4-color PNH assay for neutrophil and monocyte by flow cytometry

目的验证基于CD157的四色流式细胞检测方法(简称CD157流式法)用于阵发性睡眠性血红蛋白尿(PNH)克隆检测的可行性,并为PNH克隆检测提供一种FLAER法的替代方法。方法评估CD157流式法检测PNH克隆的批内和批间精密度。分别采用CD157流式法、FLAER法和CD55/CD59流式法对45份含不同克隆大小的PNH患者样本进行测定,并比较3种方法诊断PNH的效能。结果CD157流式法检测中性粒细胞PNH克隆和单核细胞PNH克隆的批内、批间精密度[以变异系数(CV)表示]分别为0.07%~3.01%、0.19%~7.19%和0.28%~4.95%、0.27%~7.98%。CD157流式法与FLAER法中性粒细胞及单核细胞PNH克隆的检测结果差异均无统计学意义(P>0.05)。2种方法的相关性良好(r2>0.99),一致性良好(中性粒细胞和单核细胞PNH克隆的平均偏差分别为0.13%、0.07%)。CD157流式法和FLAER法诊断PNH的敏感性(均为100.0%)高于CD55/CD59流式法(94.0%)。结论CD157流式法的批内检测性能良好,FLAER法和非FLAER法间具有良好的相关性和一致性;在不使用FLAER的情况下,或可作为FLAER法的替代方法。

Objective To validate the feasibility of CD157-based 4-color flow cytometry for paroxysmal nocturnal hemoglobinuria(PNH)clone determination,and to propose a non-FLAER alternative for PNH clone determination.Methods The intra-and inter-assay precisions of the CD157 flow cytometry for PNH clone determination were validated.CD157 flow cytometry,FLAER and CD55/CD59 methods were used to determine 45 samples of PNH patients with different clone sizes.The efficiency of the 3 methods for PNH diagnosis was evaluated..Results The intra-and inter-assay precisions[coefficients of variation(CV)]for neutrophil and monocyte were 0.07%-3.01%,0.19%-7.19%,0.28%-4.95%and 0.27%-7.98%,respectively.There was no statistical significance between CD157 flow cytometry and FLAER for neutrophil and monocyte PNH clone determination(P>0.05).There was a good correlation between the 2 methods(r2>0.99)and a good consistency,and the average deviations for neutrophil and monocyte PNH clone determination were 0.13%and 0.07%.The sensitivities of CD157 flow cytometry and FLAER in predicting PNH(100.0%)were higher than that of CD55/CD59 method(94.0%).Conclusions CD157 flow cytometry has good performance,which can be used as a FLAER alternative for PNH clone determination.