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

目的 建立并标准化流式细胞术外周血淋巴细胞亚群绝对含量的测定方法,探讨单平台和双平台法检测的特点与适用性.方法 以三色流式细胞术,采用单平台和双平台法定量测定194例健康人外周血T细胞、B细胞、NK细胞亚群,并调查两种方法检测结果的精密性.结果 单平台法健康人外周血总T细胞、T4细胞、T8细胞、T4/T8比值、DN-T细胞、DP-T细胞、B细胞和NK细胞的绝对含量依次为(1.69±1.10)、(1.10±0.74)、(0.73±0.60)、(1.51±0.62)、(0.15±0.10)、(0.02±0.03)、(0.36±0.32) 和(0.37±0.41)×109/L;双平台法结果依次为(1.87±0.70)、(1.07±0.41)、(0.74±0.31)、(1.45±0.38)、(0.15±0.07)、(0.02±0.01)、(0.36±0.16)和(0.36±0.16)×109/L,两种方法检测结果之间差异无统计学意义(t检验,P＞0.05).双平台法检测结果的变异系数在4.5%～9.0%,均低于单平台法检测结果的变异系数值(F检验,P＜0.01).结论 流式淋巴细胞亚群绝对计数的单平台法和双平台法均可用于临床检验,且单平台法检测结果的精密度更高,稳定性更好.

Experimental study on the quantitative assay of PBL subgroup by flow cytometric single-platform method and dual-platform method

Objective To establish and standardize the quantitative assay of peripheral blood lymphocytes (PBL) subgroup by flow cytometry,and investigate the characteristic and the applicability of single-platform method and dual-platform method.Methods 3-color flow cytometric single-platform method and dual-platform method were respectively used to detect quantity of each PBL subgroup from 194 healthy persons and compared in result precision.Results The levels of T cell,T4 cell,T8 cell,DN-T cell,DP-T cell,B cell and NK cell were (1.69±1.10),(1.10±0.74),(0.73±0.60),(1.51±0.62),(0.15±0.10),(0.02±0.03),(0.36±0.32) and (0.37±0.41)×109/L respectively by single-platform method,but (1.87±0.70),(1.07±0.41),(0.74±0.31),(1.45±0.38),(0.15±0.07),(0.02±0.01),(0.36±0.16) and (0.36±0.16)×109/L by dual-platform method.There was no difference between the results from above two methods by t test.The CV of dual-platform method was from 4.5% to 9.0%,and lower than single-platform method by F test.Conclusion The dual-platform method might be more precise and more stable than single-platform method,but both the two methods could be used as quantitative assay of PBL subgroup.