Flow cytometry detection of platelets autoantibodies in children with idiopathic thrombocytopenic purpura

Indian J Hematol Blood Transfus. 2009 Sep;25(3):96-103. doi: 10.1007/s12288-009-0028-0. Epub 2009 Nov 12.

Abstract

Immune thrombocytopenic purpura is an acquired disorder, in which accelerated platelet consumption is due to platelet autoantibodies. The aim of this study was to investigate the clinical value of platelet autoantibodies assay in children with ITP and to evaluate flow cytometry in the detection of platelet autoantibodies in comparison with monoclonal antibody specific immobilization of platelet antigen (MAIPA) assay. We measured platelet autoantibodies by flow cytometry and MAIPA in 18 children with ITP (6 acute, 7 chronic and 5 in remission), in addition to 5 healthy children with matched age and sex as a control group. Significant elevation of platelet-associated immunoglobulin G (PAIgG), PAIgM and PAIgA was demonstrated in children with acute ITP compared to controls and children with chronic ITP (P < 0.05). There was significant elevation of PAIgG and PAIgM in children with acute ITP compared to children with ITP in remission (P < 0.05). There was significant negative correlation between platelet count and PAIgG levels in ITP children (r = -0.717; P = 0.001). Flow cytometry found PAIgG in 94.4% of ITP children. MAIPA has detected platelet specific IgG autoantibodies in 83.3% of ITP children. ROC analysis revealed sensitivity of 94%, specificity of 57% with overall accuracy of 83% for detection of PAIgG by flow cytometry compared to MAIPA.

Conclusions: Platelet autoantibodies testing in ITP can discriminate acute from chronic forms of the disease and is helpful in follow up of patients. Determination of PAIgM in combination of PAIgG could be of interest in the investigation of ITP. Flow cytometry is a sensitive method of detection of platelet autoantibodies that could be used in screening of suspected ITP and should be followed by MAIPA assay in positive cases to establish the diagnosis.

Keywords: Flow cytometry; ITP; MAIPA.