流式细胞术检测RhD(-)血液中微量RhD(+)红细胞的研究  

Study on the detection of RhD(+) red blood cells mixed in D-negative blood by flow cytometry

作　　者：周英[1] 陈雪[1] 李健[1] 吕文彬[1] 王乃红[1] 兰炯采[2] 方军[3] 

机构地区：[1]成都市血液中心,四川成都610041 [2]南方医科大学医院 [3]成都市妇产科医院

出　　处：《中国输血杂志》2008年第5期346-348,共3页Chinese Journal of Blood Transfusion

摘　　要：目的探讨建立流式细胞术(FCM)检测微量RhD(+)红细胞的试验方法。方法选取标准的RhD(+)和RhD(-)细胞按不同比例混合,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的最佳使用剂量。在此实验条件下,作FCM检测值与实际值的相关性分析和FCM的检测重复性分析。结果抗-D的最佳使用剂量是1∶4稀释,50μl/1×106细胞;当RhD(+)细胞占RhD(-)细胞比例范围为2.5%—0.312%时,FCM测定比例与已知实际比例的相关系数(r)=0.987。RhD(+)细胞比例分别为10%、2.5%的同一测定管各重复测定10次,变异系数分别3.4%、4.9%。结论FCM检测微量RhD(+)细胞具有较好的准确度和重复性,可应用于临床检测RHD(-)患者体内的微量RhD(+)细胞。Objective To establish a method for detection of RhD（ ＋ ） red blood cells mixed in D-negative blood by flow cytometry（FCM）. Method RhD（ ＋ ） and RhD（ - ） RBCs were mixed according to predefined ratios. Cells were indirectly labeled, with IgG anti-D labeled as the first antibody, and FITC-anti-IgG F（ab＇）2 as the second antibody. The percentage of RhD（ ＋ ） RBCs was determined by FCM, and the best dosage ofIgG anti-D was also defined. The ratio of red cells in the two groups, measured by FCM, was compared with the actual ratio. The consistency of method was also evaluated. Results The effective dosage of IgG anti-D was 1：4, and 50μl/1×10^5 cells. When the actual percentages of RhD（＋） cell among RhD（-） cells were 2.5%-0.312%, the correlation coefficient between the percentages meas- ured by FCM and the actual percentages was 0.987. The same tubes, containing 10% and 2.5% RhD（＋） RBCs, were each tested for 10 times, and their coefficient of variation were 3.4%, and 4.9%, respectively. Conclusion The method of quantifying the RhD（＋） RBCs in D-negative blood by FCH is feasible and repeatable, which deserves a further clinical application.

关 键 词：红细胞 RhD(+) 流式细胞术 微量检测 

分 类 号：R457.1[医药卫生—治疗学]