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  • Title: Relative sensitivity of direct antiglobulin test, antibody's elution and flow cytometry in the serologic diagnosis of immune hemolytic transfusion reactions.
    Author: Alvarez A, Rives S, Montoto S, Sanz C, Pereira A.
    Journal: Haematologica; 2000 Feb; 85(2):186-8. PubMed ID: 10681727.
    Abstract:
    BACKGROUND AND OBJECTIVE: Current immunohematology practice dictates that serologic diagnosis of immune hemolytic transfusion reactions (IHTR) is based on the finding of a positive post-transfusion direct antiglobulin test (DAT). However, since DAT may fail to detect antibody-coated cells when they constitute a minor population amid a large number of non-sensitized ones, we investigated whether antibody detection in eluates or by flow cytometry is more sensitive than DAT in this context. DESIGN AND METHODS: Ten samples of red blood cells sensitized with allo- or autoantibodies were diluted in non-sensitized red blood cells to final concentrations ranging from 10% to 0.1%. DAT, antibody detection in eluates, and immunofluorescence by flow cytometry were performed on each mixture. RESULTS: DAT failed to detect sensitized cells in all but two cases in that only the 10% dilution yielded a positive DAT. Antibody detection in eluates and by flow cytometry was able to detect up to 1% sensitized cells in most cases. INTERPRETATION AND CONCLUSIONS: Antibody detection in eluates and by flow cytometry is more sensitive than DAT for detecting minor populations of IgG-coated cells. These techniques should be included in the routine investigation of suspected cases of IHTR.
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