选用不同的抗体组合对流式细胞术检测淋巴细胞亚群的差异研究

Analyze of the difference and clinical significance of lymphocyte subsets by flow cytometry using different combinations of antibodies

目的探讨不同的抗体组合对流式细胞术检测人外周血淋巴细胞亚群的差异及临床意义。方法随机选取江门市中心医院2012年5-12月健康体检者50例,采用双色法CD3-FITC/(CD16+56)-PE、CD3-FITC/CD19-PE分别检测NK、B淋巴细胞,四色法CD45-FITC/CD56-RD1/CD19-ECD/CD3-PC5检测NK、B淋巴细胞及四色法CD45-FITC/CD4-RD1/CD8-ECD/CD3-PC5检测T淋巴细胞,流式细胞仪直接计淋巴细胞的百分数,并用统计学方法分析比较结果。结果双色法检测的NK淋巴细胞的百分数显著高于四色法,差异有统计学意义(P<0.05),双色法检测的B淋巴细胞百分数与四色法检测的结果差异无统计学意义(P>0.05),两种方法检测的总淋巴细胞数差异有统计学意义(P<0.05)。结论应有流式细胞术检测人外周NK细胞亚群的检测应该选用双色标记的CD3-FITC/(CD16+56)-PE抗体。

Objective To explore the differences of human peripheral blood lymphocyte subsets by flow cytometry using different combinations of antibodies. Method The T cells, B cells, NK cells of 50 cases were analyzed by flow cytometry using the combinations of different antibodies：CD45-FITC/CD4-RD1/CD8-ECD/CD3-PC5,CD3-FITC/（CD16＋56）-PE, CD3-FITC/CD19-PE,and CD45-FITC/CD56-RD1/CD19-ECD/CD3-PC5. Results The NK lymphocyte percentage of two- color method detection was significantly higher than that of the four color method, and the difference was statistically significant（P〈0.05）. The B lymphocyte percentage was not different between two-color method and the four color method （P〉0.05）; the total number of lymphocytes were significantly different between two-color method and the four color method（P〈0.05）. Conclusion The CD3-FITC/（CD16＋56）-PE antibodies should be chosen to detect peripheral NK cells by flow cytometry.