流式细胞术检测胎儿红细胞的影响因素探讨

目的 探讨流式细胞术检测胎儿红细胞的影响因素。方法45名健康产妇脐血样本分别用同血型健康人外周血调整为5种浓度，使用抗胎儿血红蛋白（HbF）单克隆抗体，用直接标记和间接标记2种方法分别测定HbF阳性细胞比例．将样本放置24h、48h和7211后再次测定。结果以Fetal Cell Count TM Kit Ⅱ双色标记碳酸酐酶和HbF抗体[碳酸酐酶-异硫氰酸荧光素（CA-FITC）、HbF-藻红蛋白（PE）]为标准，5种浓度的直接标记结果均高于间接标记（P〈0．05），阳性细胞数低时差异有统计学意义（P〈0．01）。样本放置48h直接标记结果未见明显下降，间接标记结果下降明显，放置72h直接标记结果也有明显下降（P〈0．05）。结论利用流式细胞仪检测母血中胎儿红细胞含量受到标记方法、样本放置时间等因素的影响。较好的方法是采用直接标记法，染色完毕立即检测，最好不要超过3d，样本必须避光保存。

Study on the influencing factors in the detection of fetal red cells by flow cytometry

Objective To investigate the factors that may influence the detection of fetal red cells by flow cytometry. Methods Umbilical bloods from 45 healthy lying-in wemen were adjusted to five different concentrations with peripheral bloods of the same blood type. Monoclonal antibody against the fetal hemoglobin （HbF） was used to detect the quantity of fetal red cells in maternal blood by direct and indirect immunofluorescent stainings, and detect the sample after 24,48 and 72 hours again. Results Using the duel-staining of Fetal Cell Count TM Kit Ⅱ and HbF antibody （ CA- FITC. HbF-PE） as a standard method, the results of direct staining were higher than that of indirect staining, and this was more significant when the quantity was low （P 〈0. 01 ）. The results of direct staining descented a little when testing the sample after 24 and 48 hours, but they descented obviously after 72 hours （P 〈 0. 05）. The results of indirect staining descented rapidly after 48 hours （ P 〈 0.05）. Conclusions The detection of fetal red cells by flow cytometry is influenced by staining methods and sample standing-time. The recommendable method is staining with direct immunofluorescence and testing ilnmediately after staining or within three days avoiding direct light.