单平台与双平台流式细胞术T细胞亚群绝对计数的比较

doi:10.3969/j.issn.1673-8640.2016.07.012

摘要/Abstract

摘要： 目的

寻找基于CYTOMICS FC500流式细胞仪（简称CYTOMICS FC500）的相对稳定、经济的外周血T细胞亚群绝对计数方法。

方法

分别用基于CYTOMICS FC500的双平台和单平台流式细胞术（FCM）同时检测280例患者外周血T细胞亚群的绝对值,比较2种方法结果的相关性和一致性;评价高值和低值样本用2种方法检测的批内和批间精密度。

结果

双平台与单平台FCM T细胞亚群绝对计数结果的相关性好（r≥0.984）,但存在明显差异（P<0.01）,双平台FCM检测的CD4+CD8-（CD4+）T细胞绝对值比单平台FCM平均低36.5（-71.8,114.7）个/μL,差异百分比为8.7%（-13.8%,31.2%）;双平台FCM检测的CD4-CD8+（CD8+）T细胞绝对值比单平台FCM平均低41.4（-84.4,167.2）个/μL,差异百分比为8.7%（-13.9%,31.3%）;双平台FCM检测的CD4-CD8-T细胞（DNT）和CD4+CD8+T细胞（DPT）绝对值也均低于单平台FCM。双平台FCM检测CD4+T细胞、CD8+T细胞、DNT、DPT绝对值高值和低值的批内变异系数（CV）分别为3.5%和4.2%、3.8%和3.4%、6.1%和7.6%、9.5%和18.7%,批间CV分别为5.1%和5.9%、5.5%和5.1%、7.7%和9.6%、12.2%和24.6%,低于单平台FCM检测各T细胞亚群绝对值高值和低值的批内CV（4.0%和4.8%、4.2%和4.0%、6.7%和8.6%、11.1%和21.4%）和批间CV（6.3%和7.5%、7.0%和6.8%、8.5%和10.3%、13.6%和26.4%）。

结论

基于CYTOMICS FC500的双平台和单平台FCM外周血T细胞亚群绝对计数结果差异较大,不宜在同一时间段内交替使用,其中双平台FCM精密度更高,且能节约成本,可优先选择。

关键词: 流式细胞术, T细胞亚群, 绝对计数, 单平台, 双平台

Abstract:

Objective To find out a stable and economical method based on CYTOMICS FC500 for T cell subset absolute counts in peripheral blood. Methods T cell subset absolute counts in 280 patients' peripheral blood were determined by single-platform and double-platform CYTOMICS FC500 flow cytometry（FCM）,and the consistency and correlation of the 2 methods were analyzed. The within-run and between-run precisions of the 2 methods in high-level and low-level samples were evaluated. Results The double-platform FCM correlated well with single-platform FCM for T cell subset absolute counts（r≥0.984）,but T cell subset absolute counts differed between the 2 methods（P<0.01）. The mean absolute count of double-platform FCM for CD4⁺CD8-（CD4+）T cell was lower than that of single-platform FCM by 36.5 （-71.8,114.7）cells/μL,and the mean bias percentage was 8.7%（-13.8%,31.2%）. The mean absolute count of double-platform FCM for CD4-CD8⁺（CD8+）T cell was lower than that of single-platform FCM by 41.4 （-84.4,167.2）cells/μL,and the mean bias percentage was 8.7%（-13.9%,31.3%）. The absolute counts of CD4-CD8- T cell（DNT） and CD4+CD8+ T cell（DPT） obtained by double-platform FCM were lower than those by single-platform FCM. The within-run coefficients of variation（CV） of double-platform FCM for CD4+ T cell,CD8+ T cell,DNT cell and DPT cell in high-level and low-level samples were 3.5% and 4.2%,3.8% and 3.4%,6.1% and 7.6%,and 9.5% and 18.7%. The between-run CV were 5.1% and 5.9%,5.5% and 5.1%,7.7% and 9.6%,and 12.2% and 24.6%,which were lower than those of single-platform FCM. The within-run CV of single-platform were 4.0% and 4.8%,4.2% and 4.0%,6.7% and 8.6%,and 11.1% and 21.4%,and the between-run CV were 6.3% and 7.5%,7.0% and 6.8%,8.5% and 10.3%,and 13.6% and 26.4%. Conclusions The differences of T cell subset absolute counts between double-platform and single-platform FCM based on CYTOMICS FC500 are too big to use the 2 methods alternately. Furthermore,double-platform FCM is precise and economical,which is a good choice.

Key words: Flow cytometry, T cell subsets, Absolute counts, Single-platform, Double-platform

中图分类号:

R446.61

胡晓蕾, 周灵玲, 赵志钢. 单平台与双平台流式细胞术T细胞亚群绝对计数的比较[J]. 检验医学, 2016, 31(7): 593-598.

HU Xiaolei, ZHOU Lingling, ZHAO Zhigang. Comparison of T cell subset absolute counts between single-platform and double-platform flow cytometry[J]. Laboratory Medicine, 2016, 31(7): 593-598.

图/表 5

参考文献 9

1	SHI YY,LI ZY,ZHAO MH,et al. The CD4 lymphocyte count is a better predictor of overall infection than the total lymphocyte count in ANCA associated vasculitis under a corticosteroid and cyclophosphamide regimen:a retrospective cohort[J]. Medicine(Baltimore),2015,94(18):e843.
2	WADE D,DANEAU G,ABOUD S,et al. WHO multicenter evaluation of FACSCount CD4 and Pima CD4 T-cell count systems:instrument performance and misclassification of HIV-infected patients[J]. J Acquir immune Defic Syndr,2014,66(5):e98-e107.
3	PATTANAPANYASAT K,PHUANG-NGERN Y,SUKAPIROM K,et al. Comparison of 5 flow cytometric immunophenotyping systems for absolute CD4+ T-lymphocyte counts in HIV-1-infected patients living in resource-limited settings[J]. J Acquir Immune Defic Syndr,2008,49(4):339-347.
4	KOYALTA D,JENABIAN MA,NADJIOUROUM N,et al. Single-platform,volumetric,CD45-assisted pan-leucogating flow cytometry for CD4 T lymphocytes monitoring of HIV infection according to the WHO recommendations for resource-constrained settings[J]. BMC Res Notes,2013,6:169.
5	REIMANN KA,O'GORMAN MR,SPRITZLER J,et al. Multisite comparison of CD4 and CD8-T lymphocyte counting by single-versus multiple-platform methodologies:evaluation of Beckman Coulter flow-count fluorospheres and the tetraONE system. The NIAID DAIDS New Technologies Evaluation Group[J]. Clin Diagn Lab Immunol,2000,7(3):344-351.
6	HULTIN LE,CHOW M,JAMIESON BD,et al. Comparison of interlaboratory variation in absolute T-cell counts by single-platform and optimized dual-platform methods[J]. Cytometry B Clin Cytom,2010,78(3):194-200.
7	吴丽娟,刘霞,赵文利. 流式单平台和双平台计数定量检测外周血淋巴细胞亚群的实验研究[J]. 国际检验医学杂志,2012,33(2):143-145.
8	WHITBY L,WHITBY A,FLETCHER M,et al. Current laboratory practices in flow cytometry for the enumeration of CD4(+) T-lymphocyte subsets[J]. Cytometry B Clin Cytom,2015,88(5):305-311.
9	王蓓丽,郭玮,潘柏申. 室温放置时间对血凝四项检测结果的影响[J]. 中华检验医学杂志,2011,34(7):595-597.

指标	例数	FCM绝对计数[M（P25,P75）,个/μL]		单平台与双平台FCM的差异
指标	例数	单平台	双平台	绝对计数（95% CI,个μ/L）	百分比（95% CI,%）
CD4+T细胞	280	449（298,707）	413（257,659）*	36.5（-71.8,114.7）	8.7（-13.8,31.2）
CD8+T细胞	280	479（286,710）	450（258,656）*	41.4（-84.4,167.2）	8.7（-13.9,31.3）
DNT	149	45（23,77）	38（21,71）*	6.0（-10.3,22.4）	10.7（-8.6,29.9）
DPT	149	7（3,12）	6（3,11）*	0.8（-1.6,3.1）	9.3（-16.9,35.5）

指标	例数	FCM绝对计数[M（P25,P75）,个/μL]		单平台与双平台FCM的差异
指标	例数	单平台	双平台	绝对计数（95% CI,个μ/L）	百分比（95% CI,%）
CD4+T细胞	280	449（298,707）	413（257,659）*	36.5（-71.8,114.7）	8.7（-13.8,31.2）
CD8+T细胞	280	479（286,710）	450（258,656）*	41.4（-84.4,167.2）	8.7（-13.9,31.3）
DNT	149	45（23,77）	38（21,71）*	6.0（-10.3,22.4）	10.7（-8.6,29.9）
DPT	149	7（3,12）	6（3,11）*	0.8（-1.6,3.1）	9.3（-16.9,35.5）

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