应用流式细胞术对自身免疫性溶血性贫血患者自身抗体掩盖下血型同种抗体检测的研究

Application of Flow Cytometry to the Detection of Blood Group Isoantibodies under the Cover of Autoantibodies in Autoimmune Hemolytic Anemia Patients

目的在自身免疫性溶血性贫血(自免溶贫)患者血样的检测中,自身抗体往往易被同种抗体掩盖,本研究试图通过对流式细胞术与传统血清学方法的比较研究,分析两者对自身抗体中混合同种抗体的检测能力差异,为自免溶贫患者输血前检测提供更敏感可靠的方法。方法随机选择使用5例人源抗-D及4例抗-E分别与经验证不具血型特异性的自身抗体按比例梯度混合,配制模拟自免溶贫患者血浆,使用凝胶卡法与流式细胞术分别对上述模拟血浆进行检测。结果三个效价组中(16,32,64),流式方法可检测出效价为自身抗体1/8~1/16的同种抗体,灵敏度均明显强于凝胶卡方法,且检出下限为凝胶卡方法的4倍(P<0.05)。结论流式细胞术较传统血清学方法对自身抗体掩盖下同种抗体的检测具有更高的灵敏度,采用该方法用于自免溶贫患者输血前检查可降低已有同种抗体介导的输血免疫风险,提高此类患者输血安全。

Objective To compare detection capabilities between flow cytometry and a conventional serological method to develop a more sensitive and reliable technique for clinical pre-transfusion detection of autoimmune hemolytic anemia patients.Methods We used the plasma samples collected from autoimmune hemolytic anemia patients simulated with human-derived autoantibodies and alloantibodies.Common clinically significant antibodies were selected,and 5 cases of anti-D and 4 anti-E were mixed with the autoantibodies in proportion to prepare autoimmune hemolytic anemia patients’plasma samples.The gel card method and flow cytometry were used to detect the above-mentioned simulated plasma,respectively.Results In 3 titer groups(16,32 and 64),the sensitivity of flow cytometry was significantly stronger than that of the gel card method,and the detection limit was 4 times that of the gel card method(P<0.05).Conclusion Compared with the traditional serological method,flow cytometry has higher sensitivity for the detection of alloantibodies under the cover of autoantibodies.Application of this method for the pre-transfusion examination of autoimmune hemolytic anemia patients can reduce blood transfusion risks mediated by the same antibody and improve the safety of blood transfusion among them.