An improved flow cytometric immunobead array to detect autoantibodies in plasma from patients with immune thrombocytopenic purpura

作者：Zhao, Yunxiao; Zhu, Mingqing; Jiang, Miao; Zuo, Bin; Wu, Qingyu; Ruan, Changgeng; He, Yang^*

来源：Clinica Chimica Acta, 2015, 438: 396-400.

DOI：10.1016/j.cca.2014.09.018

• Flow cytometry
• Immune thrombocytopenic purpure
• Immunebead array
• Platelet glycoprotein
• Thrombocytopenia

Background: Autoantibodies against platelet glycoproteins (GPs) play an important role in immune thrombocytopenic purpura (ITP). This study was to develop an improved flow cytometric immunobead array (FCIA) assay to detect platelet autoantibodies in ITP patient plasma.
Methods: Plasma samples were isolated from 71 ITP patients and 136 non-ITP controls and incubated with platelets from normal individuals. After washing, platelets were lysed and the platelet lysates were incubated with polystyrene microbeads coupled with monoclonal antibodies against human GPs IX (SZ1), Ib (SZ2), IIIa (SZ21), IIb (SZ22), and P-selectin (SZ51). Platelet GP-autoantibody complexes were detected by flow cytometty using a FITC-labeled secondary antibody.
Result Autoantibodies against platelet GPIb, GPIIb, GPIIIa, GPIX and P-selectin were detected in plasma from ITP patients, as indicated by high mean fluorescent intensity values when microbeads with antibodies SZ1, SZ2,SZ21, SZ22, and SZ51 were used. In ROC analysis, values of the area under the curve were 0.74, 0.83, 0.80, 0.79 and 0.87, respectively. Compared with the previously reported assays, this new FCIA eliminated the need of isolating platelets from ITP patients without compromising assay sensitivity and accuracy in predicting ITP.
Conclusion: This simplified FICA assay may be more suitable for ITP diagnosis in clinical laboratory settings.

• 出版日期2015-1-1
• 单位苏州大学