Paroxysmal Nocturnal Hemoglobinuria (PNH) is a rare blood disorder characterized by chronic intravascular hemolysis, thromboses in unusual sites and cytopenias related to bone marrow failure. The diagnosis is based on the Flow Cytometric (FCM) detection of peripheral blood cell clones lacking the surface molecules linked to the GPI anchor, which is altered by mutations. Consensus studies have developed standardized and robust multicolor FCM assays to disclose PNH clones among red cells, neutrophils and monocytes at a high level of sensitivity and accuracy. High-resolution procedures have been also established to detect small PNH clones at a sensitivity level of around 0.01% in red cells and neutrophils. Cell clone type and size have been put into correlation with the clinical presentations of the associated diseases, and recommendations for the clinical follow-up have been established. The recent advent of the therapeutic monoclonal antibody Eculizumab has dramatically improved both the quality of life and the life expectancy of the affected patients, further increasing the importance of an accurate FCM detection and monitoring of the clones. The technical features of the FCM diagnostic workup and the many critical aspects of the analytical process are discussed here.
Keywords: FLAER; ICCS/ESCCA guidelines; PNH; flow cytometry; monoclonal antibodies; paroxysmal nocturnal hemoglobinuria.
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