流式细胞仪全血红细胞裂解液研究

Study on Whole Blood Red Cell Lysing Reagent for Flow Cytometry

目的研制流式细胞仪全血红细胞裂解液,并与进口试剂比对效果,以期大大降低试剂的成本,使其在中国广泛应用。方法采用流式细胞术,分别用进口试剂和自制试剂制备全血样品,比较两种试剂对30例健康体检者白细胞分群的影响、淋巴细胞亚群比例的影响、淋巴细胞亚群的平均荧光强度的影响。结果自制试剂制备的样品中淋巴细胞、单核细胞和粒细胞的分布与进口试剂一致;自制试剂制备的样品,T淋巴细胞(CD3+)、B淋巴细胞(CD19+/CD3-)、辅助/诱导T淋巴细胞(CD3+CD4+CD8-)、抑制/细胞毒T淋巴细胞(CD3+CD4-CD8+)、NK细胞(CD16+56+/CD3-)的百分率与进口试剂无明显差异(P>0.05);自制试剂制备的样品,T淋巴细胞、B淋巴细胞、辅助/诱导T淋巴细胞和抑制/细胞毒T淋巴细胞的平均荧光强度与进口试剂无明显差异(P>0.05),但NK细胞的平均荧光强度弱于进口试剂(P<0.05)。结论与进口试剂比对的结果表明,自制试剂对白细胞的分群、淋巴细胞亚群的百分率、淋巴细胞表面分子CD3、CD4、CD8、CD19的表达量,均达到了进口试剂的效果,而成本大大降低。

Objective To study and develop the whole blood red cell lysing reagents in order to replace the commercial kit and reduce the cost. Methods Flow cytometry was used to compare the home - made red cell lysing reagents with the commercial kit on the separation of white cells into three groups, on the ratio of lymphocyte subsets and on the mean fluorescence intensity （MFI） of lymphocyte subsets. Results Compared with the commercial kit, the home - made lysing reagents had no significant difference on the separation of white cells into three groups, on the ratio of lymphocyte subsets and on the MFI of T lymphocytes, B lymphocytes , Helper - Inducer T - lymphocytes and Suppressor - Cytotoxic T - lymphocytes. Conclusions The home - made lysing reagents had similar effects as the com- mercial kit on the separation of white cells, on the ratio of lymphocyte subsets and on the MFI of T lymphocytes, B lymphocytes, Helper -Inducer T- Lymphocytes and Suppressor- Cytotoxic T- lymphocytes, but the cost is much lower.