流式细胞术在感染鼠疟原虫活红细胞检测中的应用

Application of Flow Cytometry in the Detection of Mouse Malaria Parasites in Living Cells

采集感染伯氏疟原虫的ICR小鼠外周血,未经固定的红细胞用Vybrant DyeCycle Green染料和抗小鼠CD71 PE荧光抗体进行双染色(实验组)。以健康小鼠外周血作阴性对照。分别在染色后0、30和60 min利用流式细胞仪进行检测,比较各次结果的一致性。结果显示,阴性对照(健康小鼠血样)与空白对照(感染疟原虫鼠血未作染色)未检出感染红细胞的疟原虫,实验组可检出感染红细胞的疟原虫。组内相关系数(ICC)检验结果显示,3个时间点检测结果的一致性较好(ICC=0.999,P<0.01)。表明未经固定的红细胞用Vybrant DyeCycle Green染料和抗小鼠CD71 PE荧光抗体染色经流式细胞术检测可区分感染疟原虫红细胞。采集的红细胞在一定时间内未作固定对检测结果无明显影响。

Peripheral blood samples were obtained from Plasmodium berghei-infected mice, and stained with Vybrant DyeCycle Green and anti-mouse CD71 PE. With normal mouse as negative control, the blood samples were tested by flow cytometry at 0, 30, and 60 min after staining. There was no positive signal in the erythrocytes from negative control and the unstained erythrocytes from the infected mouse. However, the positive signal was detected in the stained crythrocytes from the infected mouse. The results showed an adequate consistency by intra-class correlation coefficient （ICC） test （ICC--0.999, P〈0.01）. Flow cytometry with the use of Vybrant DyeCycle Green and anti-mouse CD71 PE in staining living cells can be used for the detection of malaria parasite-infected blood. In a given period, there is no negative impact on the detection of unfixed cells.