Davis B H, Bigelow N C, Koepke J A, Borowitz M J, Houwen B, Jacobberger J W, Pierre R V, Corash L, Ault K A, Batjer J D
Department of Pathology, Dartmouth Hitchcock Medical Center, Hanover, New Hampshire.
Am J Clin Pathol. 1994 Oct;102(4):468-77. doi: 10.1093/ajcp/102.4.468.
Reticulocyte analysis by flow cytometry offers precision and sensitivity greater than those of conventional morphologic methods and permits derivation of a reticulocyte maturity index. However, interlaboratory variability has not yet been reported. The authors analyzed 310 samples at eight sites using 11 instruments over a 4-month period to examine intermethod and interlaboratory variabilities. Stains included thiazole orange, ethidium bromide, and auramine O. Instruments included models by Coulter, Becton Dickinson, TOA Medical Electronics, and Ortho Diagnostics. The coefficient of variation (CV) among all sites and methods on these samples varied as a function of the reticulocyte percentage, ranging from a mean CV of 69% for samples with < .5% reticulocytes to 24.1% for those with > 2.5% reticulocytes. The best performance was observed with the TOA R-1000 dedicated reticulocyte analyzers, with a mean CV of 18.4% for samples with < .5% reticulocytes and 4.6% for samples with > 2.5% reticulocytes. The reticulocyte maturity index showed comparable intersite precision, with a mean CV of 16% for samples with > 2.5% reticulocytes with multipurpose flow cytometers and a mean CV of 7.3% with the TOA R-1000 instruments. Interclass correlations among all sites ranged from .79 to .99 for the reticulocyte counts and .41 to .88 for the reticulocyte maturity index. The authors conclude that flow cytometric reticulocyte analysis is more precise than manual reticulocyte analysis. With greater automation of this methodology, further interlaboratory standardization of reticulocyte counts and the reticulocyte maturity index can be achieved.
通过流式细胞术进行网织红细胞分析，其精密度和灵敏度高于传统形态学方法，并且能够得出网织红细胞成熟指数。然而，尚未有关于实验室间变异性的报道。作者在4个月的时间里，使用11台仪器在8个地点分析了310份样本，以检验方法间和实验室间的变异性。所使用的染料包括噻唑橙、溴化乙锭和金胺O。仪器包括库尔特、贝克曼库尔特、东亚医疗电子和奥多诊断公司的型号。这些样本在所有地点和方法之间的变异系数（CV）随网织红细胞百分比而变化，网织红细胞<0.5%的样本平均CV为69%，而网织红细胞>2.5%的样本平均CV为24.1%。东亚医疗电子R - 1000专用网织红细胞分析仪表现最佳，网织红细胞<0.5%的样本平均CV为18.4%，网织红细胞>2.5%的样本平均CV为4.6%。网织红细胞成熟指数显示出类似的地点间精密度，使用多功能流式细胞仪检测网织红细胞>2.5%的样本平均CV为16%，使用东亚医疗电子R - 1000仪器时平均CV为7.3%。所有地点之间网织红细胞计数的组内相关性在0.79至0.99之间，网织红细胞成熟指数的组内相关性在0.41至0.88之间。作者得出结论，流式细胞术网织红细胞分析比手工网织红细胞分析更精确。随着该方法自动化程度的提高，可以实现网织红细胞计数和网织红细胞成熟指数在实验室间的进一步标准化。
