利用多指标流式细胞术和外周血标本进行白血病免疫分型的研究

目的:利用CD45/SCC组合及设门技术的多参数流式细胞术,探讨采用外周血标本在白血病免疫分型中的技术要点和资料分析中的注意事项。方法:检测标本以CD45标记,构建CD45/SSC散点图,采用一组针对不同白细胞系列的单克隆抗体进行三色或双色荧光染色,用设门技术对白血病细胞群体加以限定,进行免疫表型分析。结果:113例急性白血病患者中106例在CD45/SSC散点图上出现可供分析白血病细胞独立群体,占分析标本的94%。描述了不同白血病免疫分型的特点。讨论了运用多参数FCM白血病免疫分型的技术要点和应注意的事项。结论:多数白血病标本在CD45/SCC散点图上可出现白血病细胞群体,对白血病细胞群体设门进行分析,可迅速而可靠地获取免疫分型结果。

Immunophenotyping of leukemia with multi-parametric flow cytometry combined with CD45/SSC pattern and gating strategy using peripheral blood specimens

Objective:To explore the technical key points in data analysis of leukemia im-munophenotyping with multi-parametric flow cytometry using peripheral blood specimens.A combination pattern of CD45/side scatter(SSC)dot plot and gating strategy is applied in both data collection and analysis.Methods :Test samples are stained with monoclonal antibodies against D45and other surface markers relevant to different WBC lineages,CD45/SSC dot plot was established and leukemic cell popula-tion,once displayed,gated for immunophenotyping analysis.Results:Specimens from113patients with a-cute leukemia were analysed.A distinguishable leukemic cell population was displayed on the dot plot in106tests,accounting for 94%of the tested specimens.The characteristics of immunophenotyping in various types of leukemia were described.The important concerns in technology and data analysis were discuussed Conclusion:Leukemic cell population in the majority of patients was displayed on CD45/SSC plot.Gat-ing of leukemic cell population and analyzing of gated cell population are essential for an accurrate im-munophenotyping of leukemia.