联合分析血小板特异性抗体和瞬逝生物传感器技术检测血小板抗体的方法比较

目的 应用联合分析血小板特异性抗体(specificp platelet antibodies,SASPA)和瞬逝生物传感器技术(evanescent biosensor technology,EVA)分别检测血小板特异性同种抗体和自身抗体,对两者进行方法学比较.方法 收集94例存在HPA-1a、HPA-5b、HLA-I和自身抗体的患者血浆标本,制备血小板膜糖蛋白-血小板特异性抗体复合物,分别用SASPA和EVA方法对复合物进行检测,对SASPA检测所得平均荧光强度值(MFI)和EVA检测所得斜率值(Slope)作统计学分析和比较.结果 SASPA方法对HPA-1a、HPA-5b、HLA-I和自身抗体检测的敏感度分别为74.1％、86.9％、86.9％和40.9％,EVA方法的敏感度分别为88.9％、78.3％、60.9％和72.7％.人类血小板抗原(HPA)-1a抗体检测中,MFI与Slope表现为强相关性,相关性系数为r=O.95(P ＜0.05).结论 SASPA和EVA技术对传统的单克隆抗体特异性血小板抗原固定实验(MAIPA)作了继承和改良,SASPA方法可以用极少量的血小板同时检测多种血小板特异性抗体,提高了检测效率;EVA方法将血小板裂解后的实验时间缩短为10 min,极大地简化了操作过程,减少了检测时间.SASPA和EVA检测方法的建立对临床血小板免疫相关疾病的诊断和血小板输血领域的发展具有深远意义.

Evaluation of simultaneous analysis of specific platelet antibodies and evanescent biosensor technology technology in the identification of platelet antibodies

Objective To identify platelet allo-antibodies and auto-antibodies by simultaneous analysis of specific platelet antibodies (SASPA) and evanescent biosensor technology (EVA).Method Sera from 94 patients with HPA-1 a,HPA-5b,HLA-I antibody and auto-antibodies were collected.The glycoprotein (GP)-spe-cific platelet antibodies complexes were prepared.SASPA and EVA were performed to detect the specific platelet antibodies.The mean fluorescence intensity (MFI) and Slope values were compared and analyzed.Results For the detection of HPA-1 a,HPA-5b,HLA-I antibodies and auto-antibodies by SASPA,the sensitivity was 74.1％,86.9％,86.9％ and 40.9％,respectively.For the detection of the antibodies by EVA,the sensitivity was 88.9％,78.3％,60.9％ and 72.7％,respectively.A strong correlation between MFI and Slope values was observed in HPA-1 a detection (r =0.95,P ＜ 0.05).Conclusions SASPA and EVA are both modified methods based on monoclonal antibody specific immobilization of platelet antigen (MAIPA).SASPA proves to be a rapid assay that is available to detect several platelet-specific antibodies simultaneously with little platelets.EVA permits simple detection of platelet-specific antibodies and it reduces the testing time to 10 min after platelet lysis.SASPA and EVA pave the ways for the diagnosis of thrombocytopenic disorders and the development of clinical platelet transfusion.