A simple flow cytometric assay for routine paroxysmal nocturnal hemoglobinuria testing based on immature reticulocytes and granulocytes

Authors

  • Nikolaos J. Tsagarakis,

    Corresponding author
    1. Flow Cytometry Laboratory, Department of Immunology, Athens Regional General Hospital “G. Gennimatas,” Athens, Greece
    • Department of Immunology, Flow Cytometry Laboratory, Athens Regional General Hospital “G. Gennimatas,” 154 Messogion Avenue, Athens, 11527, Greece
    Search for more papers by this author
  • George Paterakis

    1. Flow Cytometry Laboratory, Department of Immunology, Athens Regional General Hospital “G. Gennimatas,” Athens, Greece
    Search for more papers by this author

  • How to cite this article: Tsagarakis NJ, Paterakis G. A simple flow cytometric assay for routine paroxysmal nocturnal hemoglobinuria testing based on immature reticulocytes and granulocytes. Cytometry Part B 2012; 82B: 259–263.

Abstract

Background:

The aim of this study was to test an easy-to-perform flow cytometric (FCM) assay for the routine investigation for diagnosis of paroxysmal nocturnal hemoglobinuria (PNH), through the simultaneous detection of PNH clones on immature reticulocytes (i-RET) and granulocytes.

Methods:

During the last 5 years, eight patients were diagnosed with PNH in our laboratory, among 90 patients prospectively studied for PNH. The determination of glycosylphosphatidylinositol (GPI) deficient cells on the erythroid lineage was made with a two-color FCM assay of CD71 and CD59, evaluating the PNH clone on i-RET. Three color combinations based on CD66b/CD16/CD45 and CD59/CD24/CD45 were used for the determination of GPI-deficient granulocytes.

Results:

In all the patients with PNH, the PNH clones determined with CD71(+)CD59(−) red blood cells (RBC) were nearly identical to the respective clones determined with CD16(dim/−)/CD66b(−) and CD59(−)/CD24(−) granulocytes, in contrast to the clones determined with CD59-deficient erythrocytes only, which were significantly lower.

Conclusions:

Our results indicate that the simultaneous assessment of the PNH clone on CD71(+)/CD59(−)i-RET and CD16(dim/−)/CD66b(−) granulocytes, could offer a reliable method of two series PNH screening, at low cost and with ease of application. © 2012 International Clinical Cytometry Society

Ancillary