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Comparison of traditional methods and mitogen-stimulated direct antiglobulin test for detection of anti-red blood cell autoimmunity

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Abstract

The diagnosis of autoimmune hemolytic anemia (AIHA) is based on a positive direct antiglobulin test (DAT), which is performed using various methods with different sensitivities. Recently, mitogen-stimulated (MS)-DAT was suggested to be able to identify latent anti-erythrocyte autoimmunity. Traditional methods (tube, microcolumn, and solid phase) and MS-DAT were compared in 54 consecutive cases of suspected AIHA, 28 idiopathic AIHA in clinical remission, and 12 difficult-to-diagnose cases of DAT-negative AIHA, and the results (all cases) were correlated with hematologic and hemolytic parameters. DAT tube was confirmed as the gold standard to diagnose AIHA since almost all positive cases showed hemolytic anemia and positive eluates; 10 out of 26 tube-negative cases were positive on microcolumn and solid phase antiglobulin tests, and 22 out of 26 using MS-DAT, although only half of them showed clear signs of hemolysis. Mitogen stimulation increased the amount of IgG bound to red blood cells in all groups; moreover, MS-DAT was the only positive test in 10 cases of AIHA, and mitogen stimulation facilitated the identification of autoantibody specificity in culture supernatants. We conclude that a battery of tests rather than a single test is useful for the diagnosis of AIHA, including MS-DAT as an additional test for selected cases, although the results have to be cautiously interpreted based on the overall clinical context.

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Acknowledgments

Our research was supported by these sponsors: Italian National Institute of Health (ISS), grant number: 526D/56; Fondazione Cà Granda Ospedale Maggiore Policlinico, grant number RC 2009 160.02.

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Authors and Affiliations

  1. U.O. Ematologia 2, Padiglione Granelli, Fondazione IRCCS Cà Granda Ospedale Maggiore Policlinico, Via F. Sforza 35, 20122, Milan, Italy

    Wilma Barcellini, Francesca Guia Imperiali, Anna Zaninoni & Alberto Zanella

  2. Centro Trasfusionale e di Immunoematologia, Fondazione IRCCS Cà Granda Ospedale Maggiore Policlinico, Milan, Italy

    Nicoletta Revelli, Maria Antonietta Villa, Maria Cristina Manera & Cinzia Paccapelo

Authors
  1. Wilma Barcellini
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  2. Nicoletta Revelli
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  3. Francesca Guia Imperiali
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  4. Maria Antonietta Villa
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  5. Maria Cristina Manera
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  6. Cinzia Paccapelo
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  7. Anna Zaninoni
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  8. Alberto Zanella
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Correspondence to Wilma Barcellini.

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Barcellini, W., Revelli, N., Imperiali, F.G. et al. Comparison of traditional methods and mitogen-stimulated direct antiglobulin test for detection of anti-red blood cell autoimmunity. Int J Hematol 91, 762–769 (2010). https://doi.org/10.1007/s12185-010-0578-9

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  • DOI: https://doi.org/10.1007/s12185-010-0578-9

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