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Updated: May 22, 2026
Assessing Retinal Microglial Phagocytic Function In Vivo Using a Flow Cytometry-based Assay
Published on: October 18, 2016
A simple flow cytometric assay for routine paroxysmal nocturnal hemoglobinuria testing based on immature
Nikolaos J Tsagarakis1, George Paterakis
1Flow Cytometry Laboratory, Department of Immunology, Athens Regional General Hospital G. Gennimatas, Athens, Greece. nikolaostsagarakis@gmail.com
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.
