Xu Xian-Guo, Liu Ying, Chen Shu, Hong Xiao-Zhen, Tao Su-Dan, Ma Kai-Rong, Lan Xiao-Fei, He Ji, Zhu Fa-Ming, Lyu Hang-Jun
Institute of Transfusion Medicine, Blood Center of Zhejiang Province;Key Laboratory of Blood Safety Research, Ministry of Health, Hangzhou 310006, Zhejiang Province, China.
Institute of Transfusion Medicine, Blood Center of Zhejiang Province;Key Laboratory of Blood Safety Research, Ministry of Health, Hangzhou 310006, Zhejiang Province, China. E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2015 Oct;23(5):1386-90. doi: 10.7534/j.issn.1009-2137.2015.05.031.
To detect platelet anti-HPA-1a and -1b antibodies using recombinant GPIIIa fragments coupled to Luminex beads.
The sensitivity of 2 techniques, monoclonal antibody specific immobilization of platelet antigen (MAIPA) and Luminex bead assay, was compared using 12 twofold-serial dilutions (from neat to 1 in 2048) of an anti-HPA-1a WHO international standard. The specificity of Luminex assay to identify anti-HPA-1a and -1b antibodies was assessed using 8 negative or positive controls and 36 blinded samples provided by WHO Platelet Workshop.
The sensitivity of MAIPA and Luminex bead assay to detect anti-HPA-1a was dilution 1/64 (i.e. 1.56 IU/ml) and far more than dilution 1/2048 (i.e. 0.049 IU/mL), respectively. The Luminex bead assay could specifically identify negative and positive controls of anti-HPA-1a and -1b. All results of 36 blinded samples by Luminex assay were accordant to reference results except one sample which contained high concentration antithetical antibody and resulted in false positive of anti-HPA-1b. Cross-reactivity was also not observed with the samples containing HLA, ABO or other platelet antibodies.
The Luminex beads coupled with recombinant GPIIIa fragments can be used to detect HPA-1 system antibodies with sufficient sensitivity and specificity, that is suitable for the detection of platelet alloantibodies in clinical alloimmune thrombocytopenia.
使用偶联至Luminex微球的重组糖蛋白IIIa片段检测血小板抗HPA-1a和-1b抗体。
使用抗HPA-1a世界卫生组织国际标准品的12个两倍系列稀释液（从原液到1/2048），比较单克隆抗体特异性血小板抗原固定法（MAIPA）和Luminex微球分析法这两种技术的灵敏度。使用世界卫生组织血小板研讨会提供的8个阴性或阳性对照以及36个盲法样本，评估Luminex分析法识别抗HPA-1a和-1b抗体的特异性。
MAIPA和Luminex微球分析法检测抗HPA-1a的灵敏度分别为稀释度1/64（即1.56 IU/ml）和远高于稀释度1/2048（即0.049 IU/mL）。Luminex微球分析法可以特异性识别抗HPA-1a和-1b的阴性和阳性对照。除一个样本含有高浓度的抗特异性抗体并导致抗HPA-1b假阳性外，Luminex分析法对36个盲法样本的所有结果均与参考结果一致。对于含有HLA、ABO或其他血小板抗体的样本，也未观察到交叉反应。
偶联重组糖蛋白IIIa片段的Luminex微球可用于检测HPA-1系统抗体，具有足够的灵敏度和特异性，适用于临床同种免疫性血小板减少症中血小板同种抗体的检测。
