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

doi:10.3969/j.issn.1673-8640.2022.04.015

Abstract

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

CLC Number:

R197.323.2

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.

Figures/Tables 6

References 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