FSC/SSC和CD45/SSC分群法流式T淋巴细胞亚群分析的实验研究

目的 建立并标准化流式细胞术外周血T淋巴细胞亚群含量测定方法,探讨CD45/SSC和FSC/SSC分群法检测的特点与适用性.方法 采用FSC/SSC分群的三色流式细胞术和CD45/SSC分群的四色流式细胞术,对194例健康人外周血T细胞亚群进行测定,并调查两种方法检测结果的精密性及临床应用的成功分群率.结果 FSC/SSC分群法健康人外周血总T细胞、T4细胞、T8细胞、T4/T8比值、DN-T细胞、DP-T细胞的百分含量依次为(69.98±5.79)%、(35.25±5.16)%、(25.08±4.34)%、(6.17±3.08)%、(0.71±0.36)%和(1.45±0.28)%;CD45/SSC分群法结果依次为(71.42±6.02)%、(35.82±5.78)%、(24.98±4.45)%、(5.98±3.11)%、(0.67±0.32)%和(1.47±0.30)%,两种方法检测结果之间差异无统计学意义(F检验,P＞0.05).精密度实验显示,FSC/SSC分群法和CD45/SSC分群法的变异系数均在4%以内,两者之间差异无统计学意义(P＞0.05).临床945份标本分析中,FSC/SSC分群法和CD45/SSC分群法的成功分群率分别为97.78%和99.83%,两者之间差异无统计学意义(P＞0.05).结论 FSC/SSC分群法和CD45/SSC分群法均可用于外周血淋巴细胞亚群含量的检测;FSC/SSC分群法成本相对低廉,上机操作简便,适合大规模临床应用开展,CD45/SSC分群法适合用于特殊标本的复查.

Experimental study on the detection of T-lymphocyte subsets in peripheral blood by flow cytometry with FSC/SSC or CD45/SSC gating

Objective To establish and standardize a method of percentage analysis of T-lymphocyte subsets in peripheral blood by flow cytometry and investigate the character and adaptivity of CD45/SSC and FSC/SSC clustering methods.Methods T-lymphocyte subsets in peripheral blood were detected by three-color flow cytometry of FSC/SSC clustering and four-color flow cytometry of CD45/SSC clustering respectively in 194 health people,then the precision and the clinic successful clustering ratio of the two methods were investigated.Results The percentage of T cell,CD4+T cell,CD8+T cell,DN-T cell,DP-T cell and CD4+T/ CD8+T,detected by FSC/SSC method,were (69.98±5.79)%,(35.25±5.16)%,(25.08±4.34)%,(6.17±3.08)%,(0.71±0.36)% and (1.45±0.28)%,and those,detected by CD45/SSC method were (71.42±6.02)%,(35.82±5.78)%,(24.98±4.45)%,(5.98±3.11)%,(0.67±0.32)% and (1.47±0.30)%.No significant difference between the results of the two methods(F test,P>0.05) was demonstrated.Coefficient of variability in both of the two methods were within 4%,no significant difference was recorded in the precision test(P>0.05).The successful clustering ratio of FSC/SSC and CD45/SSC methods were 97.78% and 99.83% respectively,and there was no statistical difference (P>0.05).Conclusion Both of FSC/SSC and CD45/SSC clustering methods could be used to detect T-lymphocyte subsets in peripheral blood.FSC/SSC method might have the advantage of low cost and simple operation,so it might be fit for large scale clinical application,and CD45/SSC method could be applied to recheck the special samples.