利用3种抗体流式分选母血中胎儿细胞的比较

目的比较单克隆抗转铁蛋白受体抗体(CD71McAb)、单克隆抗血型糖蛋白A抗体(GPAMcAb)和抗血红蛋白-ξ抗体(ξ-Hb)3种胎儿有核红细胞(FRBCs)特异性抗体在流式细胞仪上分选母血中胎儿细胞的检出率,评价分选胎儿细胞的最适方法。方法收集55例孕妇外周血,利用淋巴细胞分离液从中分离出单个核细胞层后,每例标本分为5组,分别加入不同抗体第1组CD71Mchb单用;第2组GPAMcAb单用;第3组ξ-Hb单用;第4组CD71+GPA联用;第5组CD71+GPA+ξ-Hb联用,孵育后流式细胞仪进行分选,应用PEP-PCR检测分选后的胎儿细胞的SRY基因。结果5组FNRBC的阳性细胞检出率及鉴定符合率分别为2.74%,77.14%;5.25%,65.4%;0.22%,95.7%;0.72%,98.6%;0.02%,60%。结论利用CD71McAb和GPAMcAb双重标记胎儿细胞进行分选或者利用抗ξ-Hb特异性McAb分选FRBCs是更有效的分选方法。

Comparison of isolating fetal erythroblasts from maternal blood by three monoclonal antibodies using flow cytometry

Objective:To compare the gain rates of isolating fetal erythroblasts from maternal blood by three monoclonal antibodies using flow cytometry, and to estimate the best method of isolating fetal cells.Methods:55 peripheral blood samples were collected from pregnant women.Each sample was divided into 5 groups. Different monoclonal antibodies were added respectively, and then they were analyzed on flow cytometry after incubated. PEP-PCR was used to detect the SRY gene of the isolated fetal cells.Results:The gain rates and the concordance rates of the five groups were: 2.74%, 77.14%; 5.25%, 65.4%; 0.22%, 95.7%; 0.72%, 98.6%;0.02%,60%.Conclusion:The methods of isolating fetal erythroblasts using anti-ξ-Hb McAb alone or anti-CD71 McAb and anti-GPA MoAb jointly are more effective.