SPR技术在红细胞血型同种抗体筛选与鉴定中的应用研究

Screening and Identification of Blood Group Alloantibody by Surface Plasmon Resonance Technique and Its Preliminary Application

目的:探讨利用表面等离子体共振(Surface plasman resonance,SPR)技术对红细胞血型同种抗体筛选与鉴定的可行性,为红细胞输注相容性检测提供一种新方法。方法:采用氨基偶联法在SPR芯片表面固定红细胞抗原谱,优化芯片分析条件,检测红细胞血型抗体阳性对照血清,分析SPR芯片技术的检测性能;对比研究SPR芯片技术与微柱凝集法检测129例长期输血的地中海贫血患者临床样本的一致性;同时运用SPR芯片技术对7例血型抗体阳性患者输血前进行血型抗体鉴定,选择红细胞抗原相容性血液输注;跟踪评价红细胞输注效果。结果:SPR芯片技术检测血型同种抗体的重复性、敏感性与特异性均较好;SPR技术与微柱凝集法对129例临床样本血型同种抗体筛查结果显示,2种方法无显著性差异(χ2=0.333, P> 0.05),总体一致性为97.2%;SPR技术对7例血型同种抗体阳性患者鉴定结果为:抗-E 3例、抗-M 1例、抗-C 1例、抗-Jka 1例、自身抗体1例,与微柱凝集法鉴定结果基本一致;SPR技术选择配合性输血后,6例患者红细胞输注有效,1例患者存在自身抗体输注无效。结论:SPR芯片技术筛检与鉴定红细胞血型抗体的性能与微柱凝集法总体一致,而SPR技术操作更简便快速、高通量且非标记,可满足批量红细胞输注前血型同种抗体快速筛查和鉴定的基本要求。

Objective:To investigate the feasibilily of screening and identifying the red blood cell type alloantibodies by means of surface plasman resonance(SPR)technique so as to provide a new method for detecting the transfusion compatibility of red blood cells.Methods:The RBC antigens for screening the alloantibody were fixed on the SPR chip surface by means of amino coupling method;the analysis conditions of SPR chip were optimized and then the control serum with RBC blood group antibody positive was detected;the performance of SPR chip for detection of serum was analysed;the consistance of rusults detected by SPR technique and microcolum agglutination for clinieal samples of129 thalasstmia patients with history of lone-term blood transfusion were compared;at the same time,the blood group amtibodies in 7 patients with blood group antibody positive were identified before blood transfusion by using SPR chip so as to select the RBC antigen compatible blood for transfusion;and the efficacy of RBC transfusion was followed up and evaluated.Results:The repeatability,sensitivity and specificity of SPR chip technique for detecting the blood group alloantibodies all were better.The SPR technique and microcolumn agglutination method were not significant different for screening blood group alloantibodies(χ^2 = 0.333,P>0.05),and the overall consistency was 97.2%;the results of SPR technique in 7 patients with positive blood group antibodies were as follows:3 cases with anti-E,1 case anti-M,1 case anti-C,1 case anti-Jka and 1 case autoantibody,which were consistent with the results of microcolumn agglutination tests,and the compatible red blood cells were selected for transfusion,of which the infusion of 6 cases was effective.In only 1 case the infusion was ineffective because of autoantibody.Conclusion:For screening and identification of blood group alloantibodies,the performance of SPR chip technique is equivalent to the micro-column agglutination,but the procedure of SPR technique is simpler,faster and high-throughput and label-free,which can meet the basic requirements for rapid screening and identification of blood group alloantibodies before transfusion of red blood cells.