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目的建立CD55、CD59流式细胞学检测平台,并探讨329例维吾尔、汉不同民族CD55、CD59表达差异。方法使用流式细胞仪对样本外周血CD55、CD59进行检测,对加入试剂量和抗体孵育时间进行摸索,对计量资料采用Multinomial Logistic回归分析。结果 CD55、CD59检测值呈现出随着试剂量增加而先增加后趋于平稳的趋势,在保证检测结果准确的前提下,为了节约成本,选择加入试剂量为10μl;随着孵育时间的延长,检测值呈现先增加后稍有降低的趋势,在抗体充分结合的前提下,为避免长时间对荧光造成淬灭的现象,选择抗体孵育时间为15min;通过对329例检测结果进行分析,发现CD55在维吾尔族和汉族人群中表达差异有统计学意义(P=0.044),与年龄、性别无关,而CD59的表达在不同的性别中稍有差异(P=0.044),与年龄、民族无关。结论分析结果提示膜蛋白缺失存在遗传易感性,这一新发现为PNH等一些血液病的研究提供了新的方向,为其临床治疗提供理论基础和新的思路。
Objective To establish a flow cytometry platform for detection of CD55 and CD59, and to explore the differences of CD55 and CD59 expression in 329 Uygur and Han ethnic groups. Methods Flow cytometry was used to detect CD55 and CD59 in peripheral blood of samples, and the amount of reagents and antibody incubation time were explored. The data were analyzed by Multinomial Logistic regression. Results The detection values of CD55 and CD59 showed the trend of increasing first and then becoming stable with the increase of reagent quantity. To save the cost and ensure the accurate test results, the amount of reagent added was 10μl. With the extension of incubation time, The detection value showed a slight increase at first and then decreased. Under the premise of sufficient antibody binding, the antibody incubation time was 15 min in order to avoid the phenomenon of quenching fluorescence for a long time. By analyzing 329 test results, it was found that CD55 There was a significant difference between Uygur and Han population (P = 0.044), but not with age and sex. However, CD59 expression was slightly different between different sexes (P = 0.044), but not with age and ethnicity. Conclusions The results suggest that there is genetic susceptibility to membrane protein deletion. This new discovery provides a new direction for the study of some hematological diseases such as PNH, and provides a theoretical basis and new ideas for its clinical treatment.