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

doi:10.3969/j.issn.1673-8640.2022.04.015

摘要/Abstract

摘要：

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。

关键词: 阵发性睡眠性血红蛋白尿, 中性粒细胞, 单核细胞, 流式细胞术

Abstract:

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.

Key words: Paroxysmal nocturnal hemoglobinuria, Neutrophil, Monocyte, Flow cytometry

中图分类号:

R197.323.2

洪俊, 饶永彩. 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[J]. 检验医学, 2022, 37(4): 377-381.

HONG Jun, RAO Yongcai. Establishment and evaluation of CD157-based 4-color PNH assay for neutrophil and monocyte by flow cytometry[J]. Laboratory Medicine, 2022, 37(4): 377-381.

图/表 6

参考文献 15

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PNH克隆	中性粒细胞			单核细胞
PNH克隆	x	s	CV	x	s	CV
大克隆	97.4	0.06	0.07	88.8	0.23	0.28
中等克隆	30.2	0.22	0.71	35.4	0.03	0.78
小克隆	0.39	0.02	3.01	0.61	0.02	4.95

PNH克隆	中性粒细胞			单核细胞
PNH克隆	x	s	CV	x	s	CV
大克隆	97.4	0.06	0.07	88.8	0.23	0.28
中等克隆	30.2	0.22	0.71	35.4	0.03	0.78
小克隆	0.39	0.02	3.01	0.61	0.02	4.95

PNH克隆小	中性粒细胞			单核细胞
PNH克隆小	x	s	CV	x	s	CV
大克隆	97.2	0.18	0.19	97.4	0.25	0.27
中等克隆	14.3	0.18	1.30	15.5	0.43	2.84
小克隆	0.49	0.03	7.19	0.79	0.04	7.98

PNH克隆小	中性粒细胞			单核细胞
PNH克隆小	x	s	CV	x	s	CV
大克隆	97.2	0.18	0.19	97.4	0.25	0.27
中等克隆	14.3	0.18	1.30	15.5	0.43	2.84
小克隆	0.49	0.03	7.19	0.79	0.04	7.98

PNH克隆	方案	中位数	最小值	最大值	95%可信区间
PNH克隆	方案	中位数	最小值	最大值	下限	上限
中性粒细胞	FLAER法	50.10	0.04	99.88	30.64	68.53
	CD157流式法	51.45	0.03	99.89	30.71	67.71
单核细胞	FLAER法	69.45	0.08	98.68	40.84	78.21
	CD157流式法	70.62	0.06	99.70	39.77	77.56