Use of CD157 in FLAER-based assays for high-sensitivity PNH granulocyte and PNH monocyte detection
Article first published online: 6 AUG 2013
© 2013 Clinical Cytometry Society
Cytometry Part B: Clinical Cytometry
Volume 86, Issue 1, pages 44–55, January 2014
How to Cite
How to cite this article: Use of CD157 in FLAERBased Assays for High-Sensitivity PNH Granulocyte and PNH Monocyte Detection. Cytometry Part B 2014; 86B: 44–55., , , , .
- Issue published online: 17 DEC 2013
- Article first published online: 6 AUG 2013
- Manuscript Accepted: 24 JUN 2013
- Manuscript Revised: 3 JUN 2013
- Manuscript Received: 1 MAR 2013
- high sensitivity 5-color flow;
- paroxysmal nocturnal hemoglobinuria;
Recent Flow Cytometric guidelines to detect Paroxysmal Nocturnal Hemoglobinuria (PNH) in white blood cells recommend using FLAER-based assays to detect granulocytes and monocytes lacking expression of GPI-linked structures. However national proficiency testing results continue to suggest a need for improved testing algorithms, including the need to optimize diagnostic analytes in PNH.
CD157 is another GPI-linked structure expressed on both granulocytes and monocytes and here we assess its ability to replace CD24 and CD14 in predicate 4-color granulocyte and monocyte assays respectively. We also assess a single tube, 5-color combination of FLAER, CD157, CD64, CD15, and CD45 to simultaneously detect PNH clones in granulocyte and monocyte lineages.
Delineation of PNH from normal phenotypes with 4- or 5-color CD157-based assays compared favorably with 4-color predicate methods and PNH clone size data were similar and highly correlated (R2 >0.99) with predicate values over a range (0.06%–99.8%) of samples. Both CD157-based assays exhibited similar high levels of sensitivity and low background levels in normal samples.
While CD157-based 4- and 5-color assays generated closely similar results to the predicate assays on a range of PNH and normal samples, the 5-color assay has significant advantages. Only a single 5-color WBC reagent cocktail is required to detect both PNH granulocytes and monocytes. Additionally, sample preparation and analysis time is reduced yielding significant efficiencies in technical resources and reagent costs. All 4- and 5-color reagent sets stained stabilized whole blood PNH preparations, used in external quality assurance programs. © 2013 International Clinical Cytometry Society