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FP5

PLATELETS Résumé de rapport

Project ID: QLK3-CT-2001-01169
Financé au titre de: FP5-LIFE QUALITY
Pays: Germany

SASPA

The German team developed and evaluated an improved MAIPA method based on simultaneous detection of various platelet specific immunoglobulin G (IgG) and IgM antibodies immobilized on beads and detected by flow cytometry.

Bead populations with distinct fluorescence intensities, coated with monoclonal antibodies specific for mouse heavy chain isotypes, were used for the simultaneous immobilization of platelet-GP [IIb/IIIa, Ib/IX, human leucocyte antigen (HLA) class I, or Ia/IIa, CD32, GPIV or CD109, Ib/IX, HLA class I]. In order to detect human IgG and IgM antibodies simultaneously, phycoerythrin- and fluorescein isotiocyanate-conjugated goat anti-human IgG and IgM were added. On this basis, the abundance of six distinct antibodies (three anti-GP, each with subclasses IgG and IgM) were simultaneously analysed without cross-reaction by flow cytometry.

For evaluation, sera and platelets from 169 patients with platelet-binding and/or platelet-associated antibodies were investigated. The monoclonal antibody-specific immobilization of platelet antigen (MAIPA) assay was performed in parallel as reference test. The simultaneous analysis of various platelet-specific antibodies (SASPA) assay was able to detect all platelet-specific IgG and IgM that were also recognized by MAIPA with a comparable sensitivity.

SASPA proved to be a rapid and reliable assay that required less platelet than other methods.

Reported by

UNIVERSITAT HEIDELBERG - DRK - BLUTSPENDEDIENST BADEN-WUERTTEMBERG
Universität Heidelberg - DRK - BLUTSPENDEDIENST BADEN WUERTEMBERG - Institue of Transfusion Medicine and Clinical Immunology - Friedrich Ebert Strasse 107
68167 MANNHEIM
Germany
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