流式细胞免疫珠阵列法检测免疫性血小板减少性紫癜患者血小板糖蛋白自身抗体。

Detection of autoantibodies against platelet glycoproteins in patients with immune thrombocytopenic purpura by flow cytometric immunobead array.

机构信息

Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Suzhou, China.

出版信息

Clin Chim Acta. 2013 Jan 16;415:176-80. doi: 10.1016/j.cca.2012.10.035. Epub 2012 Oct 25.

DOI:10.1016/j.cca.2012.10.035

PMID:23103637

Abstract

BACKGROUND

The goal of this study is to develop a flow cytometric immunobead array (FCIA) assay to detect platelet autoantibodies commonly present in bleeding patients with immune thrombocytopenic purpura (ITP).

METHODS

Polystyrene microbeads coated with antibodies against human platelet glycoproteins (GPs) IX (SZ1), Ib (SZ2), IIIa (SZ21), IIb (SZ22), and P-selectin (SZ51) were incubated with platelet lysate from 50 ITP patients and 86 controls. The platelet antigen-autoantibody complexes were detected by flow cytometry using an FITC-labeled antibody. The results were compared with that of a monoclonal antibody immobilization of platelet antigen (MAIPA) assay.

RESULTS

By FCIA, platelet autoantibodies against GPIb, GPIIb, GPIIIa, GPIX and P-selectin were detected in ITP patients. Mean fluorescent intensity values with antibodies SZ1, SZ2, SZ21, SZ22 and SZ51 were all higher in ITP patients than controls (p values<0.01). In ROC analysis, values of the area under the curve were 0.89, 0.82, 0.93, 0.94 and 0.95, respectively. In ITP diagnosis, the FCIA assay with these five antibodies had better sensitivity and accuracy than the MAIPA assay (96% vs. 44% in sensitivity; 80.9% vs. 64.7% in accuracy, p<0.01).

CONCLUSION

FCIA assays with multiple antibodies against platelet GPs may be used to improve the diagnosis of ITP in hospitals.

摘要

背景

本研究旨在开发一种流式细胞免疫珠阵列（FCIA）检测方法，用于检测常见于免疫性血小板减少性紫癜（ITP）出血患者的血小板自身抗体。

方法

用针对人血小板糖蛋白（GPs）IX（SZ1）、Ib（SZ2）、IIIa（SZ21）、IIb（SZ22）和 P-选择素（SZ51）的抗体包被聚苯乙烯微珠，与 50 例 ITP 患者和 86 例对照的血小板裂解物孵育。使用 FITC 标记的抗体通过流式细胞术检测血小板抗原-自身抗体复合物。将结果与单克隆抗体固定血小板抗原（MAIPA）检测进行比较。

结果

通过 FCIA，在 ITP 患者中检测到针对 GPIb、GPIIb、GPIIIa、GPIX 和 P-选择素的血小板自身抗体。与对照组相比，ITP 患者的抗体 SZ1、SZ2、SZ21、SZ22 和 SZ51 的平均荧光强度值均较高（p 值均<0.01）。在 ROC 分析中，曲线下面积的值分别为 0.89、0.82、0.93、0.94 和 0.95。在 ITP 诊断中，与 MAIPA 检测相比，该 FCIA 检测五种抗体的敏感性和准确性均更好（敏感性分别为 96%和 44%；准确性分别为 80.9%和 64.7%，均 p<0.01）。