摘要
本研究旨在建立流式细胞术(FCM)检测红细胞表面RhD抗原的实验方法,并研究不同RhD血清型红细胞表面D抗原表达的差异。选取标准的RhD(+)和RhD(-)细胞按1∶1比例混合,用间接标记法[一抗为IgG抗-D,二抗为FITC-抗-IgG F(ab′)2]染色,应用FCM检测RhD(+)细胞的比例,并确立IgG抗-D的最佳使用稀释度。采用FCM检测RhD阳性、弱D型、RhDel型、RhD阴性红细胞表面D抗原数量。结果显示,IgG抗-D单克隆抗体的最佳使用稀释度是1∶4,1×106/50μl。RhD阳性、弱D型、RhDel型、RhD阴性者RhD(+)红细胞百分率分别为(96.8±2.97)%、(79.5±9.88)%、(47.8±11.43)%、(3.7±2.96)%,红细胞表面D抗原平均荧光强度依次为33.3±6.21道、18.6±5.39道、7.10±1.17道、0.79±0.55道。结论:4种不同血清表现型的RhD抗原数量的差异有显著性,以RhD阳性的抗原数量最多,弱D型次之,RhDel型最少。
This study was purposed to establish the method of quantifying RhD antigen on red blood cells （RBC） by flow cytometry （FCM） and to explore the expression of D antigen on RBC of different RhD serotype. RhD（ ＋ ） RBCs and RhD （ - ） RBCs were mixed in 1 ： 1 ratio. Cells were stained by the indirect method （ IgG anti-D as the first antibody, FITC-anti-IgG F（ ab′ ） 2 as the second antibody）, and the ratio of RhD（ ＋ ） on RBCs was quantified by FCM. The optimal dosage of IgG anti-D was defined. Expression of RhD antigen on RBC of RhD（ ＋ ）, weak D, RhDel and RhD（-） type were detected by FCM. The results showed that optimal dilution of IgG anti-D monoclonal antibody was 1 ：4, 1 ×10^6 ceils/50 μl. The percentage of D（ ＋ ） RBC of RhD（ ＋ ）, weak D, RhDel and RhD（ - ） type were 96.8 ± 2.97 %, 79.5 ± 9.88 %, 47.8 ± 11.43 %, 3.7 ± 2.96 %, respectively. The mean fluorescence intensity （MFI） of RhD antigen expression of RhD（ ＋ ）, weak D, RhDel and RhD（ - ） type were 33.3 ±6.21Dal, 18.6 ±5.39 Dal, 7.10 ± 1.17Dal, 0.79 ± 0.55Dal, respectively. In conclusion, there are significant differences of RhD antigen expressions among RBC of different RhD serotypes. The level of antigen on RhD（ ＋ ） RBC is the highest and then weak D the next, while the leve of antigen on RhDel RBC is the lowest level.
出处
《中国实验血液学杂志》
CAS
CSCD
2008年第6期1452-1454,共3页
Journal of Experimental Hematology
