流式细胞术在人ABO血型特异性抗体检测中的应用

目的 建立以流式细胞仪检测人ABO天然血型特异性抗体的技术,并检验其效果.方法 将52份健康志愿者的血清样本与标准化人源红细胞混合,加入异硫氰基荧光素标记的羊抗人IgM或IgG二抗,用流式细胞仪检测样本中抗红细胞抗原A和B的特异性IgM及IgG抗体.检测结果与酶标仪法进行对比.结果两种方法对于血型IgM抗体水平变化趋势检测的关联性较好,二者的Spearman等级相关系数为0.730(P＜0.05),但流式细胞术的特异性、灵敏性和重复性优于酶标仪法,A型和B型血清样本(各16例)中,流式细胞术结果转为阴性的IgM抗体滴度为1∶1024～1∶512,而酶标仪法则为1∶64～1∶32.用流式细胞术在O型血清中检测到IgG抗体.结论 流式细胞术可以进行ABO血型IgM和IgG抗体的检测,效果优于传统方法,并可半定量检测抗体水平.

Abstract：

Objective To establish and validate a standard method using flow cytometry (FCM)in human ABO blood group antibody (Ab) detection. Methods Sera samples from 52 blood donors were incubated with standardize red blood cells (RBCs), and anti-A/B antibody (IgM/IgG) was measured by indirect flow cytometry after adding secondary isotype-specific fluoresce labeled Ab. The results were compared with those by using ELISA reader. Results The changes of anti-A/B IgM measured by both methods correlated well and an overall correlation coefficiency of 0.730 for IgM was obtained by Spearman's rank testing (P＜0.05). FCM showed better specificity, sensitivity and reproducibility over ELISA reader. In blood group A samples (n = 16) or B samples (n = 16) ,the antirespectively. IgG blood group Ab could only be detected in blood group O sera. Conclusion Compared with traditional methods,FCM is a more objective method to offer accurate detection of natural ABO blood group Ab (IgM/IgG) and allowed semi-quantitative measurement of antibody levels.

Application of flow cytometry to detect natural specific antibody levels in human ABO blood group

Objective To establish and validate a standard method using flow cytometry (FCM)in human ABO blood group antibody (Ab) detection. Methods Sera samples from 52 blood donors were incubated with standardize red blood cells (RBCs), and anti-A/B antibody (IgM/IgG) was measured by indirect flow cytometry after adding secondary isotype-specific fluoresce labeled Ab. The results were compared with those by using ELISA reader. Results The changes of anti-A/B IgM measured by both methods correlated well and an overall correlation coefficiency of 0.730 for IgM was obtained by Spearman's rank testing (P＜0.05). FCM showed better specificity, sensitivity and reproducibility over ELISA reader. In blood group A samples (n = 16) or B samples (n = 16) ,the antirespectively. IgG blood group Ab could only be detected in blood group O sera. Conclusion Compared with traditional methods,FCM is a more objective method to offer accurate detection of natural ABO blood group Ab (IgM/IgG) and allowed semi-quantitative measurement of antibody levels.