title: An improved flow cytometric immunobead array to detect autoantibodies in plasma from patients with immune thrombocytopenic purpura.
abstract: 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 cytometry using a FITC-labeled secondary antibody.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  This simplified FICA assay may be more suitable for ITP diagnosis in clinical laboratory settings.
authors.affiliation: MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital, the Cyrus Tang Hematology Center, and the Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China.
authors.forename: Yunxiao
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital, the Cyrus Tang Hematology Center, and the Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China.
authors.forename: Mingqing
authors.initials: M
authors.lastname: Zhu
authors.affiliation: MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital, the Cyrus Tang Hematology Center, and the Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China.
authors.forename: Miao
authors.initials: M
authors.lastname: Jiang
authors.affiliation: MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital, the Cyrus Tang Hematology Center, and the Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Zuo
authors.affiliation: MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital, the Cyrus Tang Hematology Center, and the Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China.
authors.forename: Qingyu
authors.initials: Q
authors.lastname: Wu
authors.affiliation: MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital, the Cyrus Tang Hematology Center, and the Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China.
authors.forename: Changgeng
authors.initials: C
authors.lastname: Ruan
authors.affiliation: MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital, the Cyrus Tang Hematology Center, and the Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China. Electronic address: heyang1963@163.com.
authors.forename: Yang
authors.initials: Y
authors.lastname: He
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000911:Antibodies, Monoclonal; D019540:Area Under Curve; D001323:Autoantibodies / Q000097:blood*; D001792:Blood Platelets / Q000737:chemistry / Q000276:immunology; D016022:Case-Control Studies; D002648:Child; D002675:Child, Preschool; D005260:Female; D005434:Flow Cytometry / Q000295:instrumentation / Q000592:standards*; D016650:Fluorescein-5-isothiocyanate; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation / Q000592:standards*; D008297:Male; D008863:Microspheres; D008875:Middle Aged; D010980:Platelet Membrane Glycoproteins / Q000032:analysis* / Q000276:immunology; D011137:Polystyrenes; D040081:Protein Array Analysis / Q000295:instrumentation / Q000592:standards*; D016553:Purpura, Thrombocytopenic, Idiopathic / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D012372:ROC Curve; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D010980:Platelet Membrane Glycoproteins; D011137:Polystyrenes; D016650:Fluorescein-5-isothiocyanate
keywords: Flow cytometry; Immune thrombocytopenic purpure; Immunebead array; Platelet glycoprotein; Thrombocytopenia