Comparison of T cell subset absolute counts between single-platform and double-platform flow cytometry

doi:10.3969/j.issn.1673-8640.2016.07.012

Abstract

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

CLC Number:

R446.61

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.

Figures/Tables 5

References 9

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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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