Primary experimental study on the reference method for the counting of red blood cells and white blood cells

doi:10.3969/j.issn.1673-8640.2015.06.023

Abstract

Abstract: Objective

To investigate the reference method for red blood cell (RBC) and white blood cell (WBC) counts, and to establish standardized determination platform which will provide technical support for blood cell counting trueness verification test.

Methods

The reference method was established according to relevant documents of the International Council for Standardization in Hematology (ICSH) and the standard of the People's Republic of China (WS/T 245-2005). The S-CAL cell calibrator, 5C cell control, aperture instrument concentration control, processed whole blood and fresh whole blood samples were determined on Z2 particle counter. Parameters such as within-run and between-run coefficients of variance (CV), coincidence counting correction, stability and comparability were evaluated.

Results

The method showed that within-run CV was<1%, and between-run CV met the requirements of the Clinical and Laboratory Standards Institute (CLSI) H26-A2. The results demonstrated the biases of coincidence counting correction in RBC and WBC counts were <1%. RBC and WBC of processed fresh whole blood samples were stable after storage for above 5 d. The biases of fresh whole blood RBC and WBC between Z2 particle counter and BECKMAN-COULTER ACT diff Ⅱ, BECKMAN-COULTER LH-750, SYSMEX K-4500, SYSMEX XE-2100 hematology analyzers were ≤±2% and ≤±4%.

Conclusions

The reference method for RBC and WBC counting by Z2 particle counter has been established basically. Precision, coincidence counting correction, accuracy, stability and comparability are conformed to the requirements. It provides the technical support for blood cell counting trueness verification test.

Key words: Red blood cell, White blood cell, Counting, Reference method, Z2 particle counter

CLC Number:

R446.11

XU Lei, WANG Qing, MIAO Yingbo, SONG Ying, WANG Guofei, ZHU Peichao, GUO Xiaojun. Primary experimental study on the reference method for the counting of red blood cells and white blood cells[J]. Laboratory Medicine, 2015, 30(6): 641-645.

Figures/Tables 7

项目	稀释管1	稀释管2	稀释管3	稀释管4
次级稀释(mL)	0.02+20^*	0.04+20^*	0.06+20^*	0.08+20^*
X轴设定值(%)	25.07	50.10	75.07	100.00
细胞计数(次数)	12	6	4	3

项目	测定次数	$x ?$	s	CV (%)	参考方法要求
RBC(×10¹²/L)	10	4.200	0.018	0.43	<1%
WBC(×10⁹/L)	10	8.550	0.065	0.76	<1%
浓度质控品	10	1.004	0.008	0.80	<1%

项目	测定次数	$x ?$	s	CV (%)	参考方法要求
RBC(×10¹²/L)	10	4.200	0.018	0.43	<1%
WBC(×10⁹/L)	10	8.550	0.065	0.76	<1%
浓度质控品	10	1.004	0.008	0.80	<1%

项目批号	测定天数(d)	$x ?$	极差	CLSI H26-A2 极差要求^[3]
RBC(×10¹²/L)889700	8	5.451	0.14	≤±0.15
RBC(×10¹²/L)879300	15	4.191	0.10	≤±0.15
WBC(×10⁹/L)889700	8	9.823	0.30	≤±0.3
WBC(×10⁹/L)868100	15	3.748	0.30	≤±0.3

项目批号	测定天数(d)	$x ?$	极差	CLSI H26-A2 极差要求^[3]
RBC(×10¹²/L)889700	8	5.451	0.14	≤±0.15
RBC(×10¹²/L)879300	15	4.191	0.10	≤±0.15
WBC(×10⁹/L)889700	8	9.823	0.30	≤±0.3
WBC(×10⁹/L)868100	15	3.748	0.30	≤±0.3

项目	重合计数校正值	靶值	偏倚(%)	方程式
RBC(×10¹²/L)	4.139 5	4.11	0.72	Y=-0.000 26X+4.139 5
WBC(×10⁹/L)	9.032 4	9.00	0.36	Y=0.000 71X+9.032 4

项目	处理方式	样品号	每周开瓶次数	稳定天数	目标值^*	结果分布	偏倚分布 (%)	允许偏倚 (%)
RBC(×10¹²/L)	方式1	0	2	22	5.193 6	5.093 85.248 7	-1.92~1.06	≤±2%
	方式2	3	2	13	3.740 4	3.671 93.757 6	-1.83~0.46	≤±2%
		23	2	9	4.954 0	4.868 14.868 2	-1.74~-1.74	≤±2%
		4	1	30	4.433 9	4.359 64.488 5	-1.68~1.23	≤±2%
	方式3	26	2	15	4.425 3	4.340 34.380 5	-1.921.01	≤±2%
项目	处理方式	样品号	每周开瓶次数	稳定天数	目标值^*	结果分布	偏倚分布 (%)	允许偏倚 (%)
WBC(×10⁹/L)	方式1	0	2	5	5.431	5.4955.552	-1.19~2.24	≤±4%
	方式2	3	2	8	5.690	5.4835.898	-3.65~3.65	≤±4%
		23	2	13	9.174	8.9039.458	-2.95~3.10	≤±4%
		4	1	28	5.761	5.7565.841	-0.09~1.39	≤±4%
	方式3	26	2	6	5.922	5.881	-0.69	≤±4%

仪器	RBC(×10¹²/L)				WBC(×10⁹/L)
仪器	样本数	与Z2的绝对误差	偏倚 (%)	允许偏倚 (%)	样本数	与Z2的绝对误差	偏倚 (%)	允许偏倚 (%)
Z2颗粒计数仪	1324				1320
ACT diffⅡ	16	-0.042	-0.94	≤±2%	17	-0.08	-1.36	≤±4%
LH-750	21	-0.012	-0.27	≤±2%	20	-0.11	-1.83	≤±4%
K-4500	24	0.032	0.73	≤±2%	13	-0.11	-1.93	≤±4%
XE-2100	13	0.042	0.95	≤±2%	13	0.06	1.05	≤±4%

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