根据 2012 年《高灵敏度阵发性睡眠性血红蛋白尿症检测实用指南》，流式细胞术检测阵发性睡眠性血红蛋白尿症的室内和室间变异性。
Intra- and interlaboratory variability of paroxysmal nocturnal hemoglobinuria testing by flow cytometry following the 2012 Practical Guidelines for high-sensitivity paroxysmal nocturnal hemoglobinuria testing.
机构信息
Institute of Hematology and Blood Transfusion, Prague, Czech Republic.
出版信息
Cytometry B Clin Cytom. 2013 Jul-Aug;84(4):229-36. doi: 10.1002/cyto.b.21075. Epub 2013 Jan 16.
BACKGROUND
Sutherland et al. recently published the Practical Guidelines for high-sensitivity detection of paroxysmal nocturnal hemoglobinuria (PNH) clones by flow cytometry (FCM), containing concise protocols for PNH testing.
METHODS
Using this approach, we studied the intra- and interlaboratory variability observed in a multicenter study in which fresh blood samples containing three clinically relevant PNH clone sizes within the granulocytic, monocytic, and red blood cell (RBC) populations were shipped to each participating center.
RESULTS
Coefficients of variation (CVs) for precision/reproducibility analysis ranged from 0.01%/0.02% to 0.48%/0.45% (big clone), from 0.69%/1.52% to 4.24%/5.80% (small-intermediate clone), from 1.47%/3.91% to 15.01% /17.83% (minor clone) for PNH white blood cells (WBCs) and from 0.24%/0.48% to 1.76%/1.83% (big clone), from 0.80%/1.14% to 2.39%/4.45% (small-intermediate clone), from 1.09%/3.36% to 10.54%/10.23% (minor clone) for PNH RBCs, respectively. Linear regression analysis showed excellent performance correlation between centers (r > 0.99), Wilcoxon rank test revealed no statistically significant differences for PNH granulocytes, monocytes, and RBCs (P > 0.05%), Bland-Altman analysis demonstrated good performance agreement for all target PNH clones (mean bias ranging from -1.47 to 0.71).
CONCLUSION
Our results demonstrate very good intra- and interlaboratory performance characteristics for both precision and reproducibility analyses and excellent correlation and agreement between centers for all target PNH clone sizes. Our data confirm the reliability and robustness of the recently published Practical Guidelines approach for high sensitivity PNH testing by flow cytometry and suggest that such an approach represents an excellent basis for standardization of PNH testing by flow cytometry.
背景
Sutherland 等人最近发表了通过流式细胞术（FCM）高灵敏度检测阵发性夜间血红蛋白尿（PNH）克隆的实用指南，其中包含了 PNH 检测的简明方案。
方法
我们使用这种方法，在一项多中心研究中研究了观察到的室内和实验室间变异性，该研究将新鲜血样运送到每个参与中心，这些血样包含粒细胞、单核细胞和红细胞（RBC）群体中的三个临床相关的 PNH 克隆大小。
结果
精密度/可重复性分析的变异系数（CV）范围为 0.01%/0.02%至 0.48%/0.45%（大克隆），0.69%/1.52%至 4.24%/5.80%（小-中间克隆），1.47%/3.91%至 15.01%/17.83%（小克隆），用于 PNH 白细胞（WBC），0.24%/0.48%至 1.76%/1.83%（大克隆），0.80%/1.14%至 2.39%/4.45%（小-中间克隆），1.09%/3.36%至 10.54%/10.23%（小克隆），用于 PNH RBC。线性回归分析显示中心之间具有出色的性能相关性（r>0.99），Wilcoxon 秩检验显示 PNH 粒细胞、单核细胞和 RBC 之间无统计学显著差异（P>0.05%），Bland-Altman 分析显示所有目标 PNH 克隆的性能一致性良好（平均偏差范围为-1.47 至 0.71）。
结论
我们的结果表明，无论是精密度还是可重复性分析，室内和实验室间的性能特征都非常出色，并且所有目标 PNH 克隆大小的中心之间都具有出色的相关性和一致性。我们的数据证实了最近发表的通过流式细胞术进行高灵敏度 PNH 检测的实用指南方法的可靠性和稳健性，并表明该方法是流式细胞术 PNH 检测标准化的极好基础。
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