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.
A simple flow cytometric assay for routine paroxysmal nocturnal hemoglobinuria testing based on immature reticulocytes and granulocytes†
Article first published online: 30 MAY 2012
Copyright © 2012 International Clinical Cytometry Society
Cytometry Part B: Clinical Cytometry
Volume 82B, Issue 4, pages 259–263, July 2012
How to Cite
Tsagarakis, N. J. and Paterakis, G. (2012), A simple flow cytometric assay for routine paroxysmal nocturnal hemoglobinuria testing based on immature reticulocytes and granulocytes. Cytometry, 82B: 259–263. doi: 10.1002/cyto.b.21030
- Issue published online: 21 JUN 2012
- Article first published online: 30 MAY 2012
- Accepted manuscript online: 26 APR 2012 02:57PM EST
- Manuscript Accepted: 14 APR 2012
- Manuscript Revised: 12 APR 2012
- Manuscript Received: 12 DEC 2011
- flow cytometry;
- PNH granulocytes;
- PNH diagnosis
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.
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.
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.
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