摘要
目的建立流式微球技术检测血小板特异性自身抗体方法,并对方法学及临床应用进行初步探讨。方法用包被抗血小板膜糖蛋白(GP)Ⅰb、Ⅱb、Ⅲa、Ⅱb/Ⅲa单克隆抗体的微球捕获与血小板GP结合的特异性抗体,加入FITC标记的羊抗人IgG抗体后用流式细胞仪检测分析。结果特发性血小板减少性紫癜(ITP)组4种单抗荧光强度比值与非ITP血小板减少组和正常对照组有显著性差异(P<0.01);若将ITP组患者4种单抗荧光强度比值分别大于正常对照组上限1.37、1.24、1.48和1.19判断为阳性,则流式微球技术检测血小板特异性自身抗体的敏感性为73.2%,特异性为94.3%;4种单抗联合检测总体敏感性明显高于改良间接单抗特异的血小板抗原固定试验(MAIPA)(P<0.05),且大于各单个抗体检测敏感性。结论流式微球技术可以简便、快捷地检测血小板膜糖蛋白特异性抗体,联合检测多种自身抗体可以提高检测阳性率,对于ITP的诊治具有一定的临床价值。
Objective To establish a method for detection of platelet-associated autoantibodies against platelet-specific receptors by cytometric bead assay. Methods The detection beads were coated by monoclonal antibodies against ptatelet glycoprotein I b, II b, IIIa, and II b/III a to capture platelet glycoprotein combined platelet-associated autoantibodies. Fluorescein isothiocyanate labeled polyclonal goat antihuman immunoglobulin antibody was added, and flow cytometer was used to detect bead-platelet-assoeiated autoantibodies-antihuman immunoglobulin antibody complex. Results The fluoreseene ratio of four monoelonal antibodies was significantly different （ P 〈 0.01 ） between the idiopathic thrombocytopenic purpura （ITP） patients and either the non-ITP patients or the normal controls. If the upper limit of normal control was set as cutoff value, ratios of greater than 1.37, 1.24, 1.48 and 1.19 were considered positive for the four monoclonal antibodies respectively. The flow cytometric bead assay had an overall sensitivity of 73. 17% and a specificity of 94.29%. The overall sensitivity was significantly higher （P 〈 0.05 ） than that of modified indirect MAIPA and that of using single antibody. Conclusions Platelet-assoeiated autoantibodies can he detected rapidly by flow cytometfic bead assay. Combined detection of autoantibodies can improve the positive rate. The assay would be valuable for diagnosis of ITP.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2010年第2期110-112,共3页
Chinese Journal of Clinical Laboratory Science
关键词
流式微球技术
自身抗体
糖蛋白
血小板减少
cytometric bead assay
autoantibody
platelet glycoprotein
thrombocytopenia
