细胞浓度对血型血清学试验灵敏度的影响

Effect of red blood cell concentration on the sensitivity of serological test detection

目的探索在常用血型血清学试验细胞浓度范围内,浓度的上限和下限对直接血凝试验及间接抗球试验的检测灵敏度是否有影响。方法采用单克隆Ig M及Ig G抗-D分别与1%、2%、3%、4%、5%、6%的R1R2细胞反应,通过试管法结合流式细胞术,检测抗体与不同浓度红细胞的反应效价,以及同一抗体浓度下不同浓度红细胞的抗体结合效率的差异。并采用酶联免疫吸附(ELISA)方法对单克隆抗体浓度进行定量。结果单克隆Ig M抗-D及Ig G抗-D的效价随着红细胞浓度的下降而依次降低,在常用血清学红细胞浓度范围内,2%红细胞与两类抗体的反应效价皆是5%的4倍。在相同抗体浓度下,细胞结合抗体平均荧光强度(MFI)随细胞浓度下降而升高。其中当Ig M、Ig G抗-D浓度分别为(25.50±1.1)μg/m L和(2.71±0.08)mg/m L时,2%红细胞结合Ig M抗-D及Ig G的MFI分别比5%红细胞高出104.2%和67.8%。结论在常用血型血清学试验细胞浓度范围内,血型血清学试验检测灵敏度随红细胞浓度下降而升高。

Objective To explore the effect of different red blood cell（ RBC） concentration on the sensitivity of serological antibody- antigen detection. Methods The concentration of monoclonal Ig M and Ig G anti- D were quantified by ELISA Kit. Monoclonal D antigens of RBCs were quantified by binding 1%,2%,3%,4%,5% and 6% R1R2 RBCs to D antigens. Using tube method and FCM（ flow cytometry）,antibodies＇ titers and the deviation in antibody bonding efficiency were detected at different RBC concentration. Results As the concentration of RBCs decreases,the titer of Ig M anti- D and Ig G anti- D also decreased. At the same antibody concentration,the antibody- bonding efficiency of 2% RBC was significantly higher than that of other RBC concentrations. When the concentrations of Ig M anti- D and Ig G anti- D were 25. 50 ± 1. 1ug / ml and 2. 71 ± 0. 08 mg / ml,the median fluorescence intensities（ MFI） of attached antibodies at 2% RBC concentration were 104. 2% and 67. 8% higher than those at 5% RBC concentration,respectively. Conclusion In routine serological testing,the sensitivity of serological detection increases as the concentration of RBC decreases.