title: Use of a FLAER-based WBC assay in the primary screening of PNH clones.
abstract: Diagnosis of paroxysmal nocturnal hemoglobinuria (PNH) with flow cytometry traditionally involves the analysis of CD55 and CD59 on RBCs and neutrophils. However, the ability to accurately detect PNH RBCs is compromised by prior hemolysis and/or transfused RBCs. Patients with aplastic anemia (AA) and myelodysplastic syndrome (MDS) can also produce PNH clones. We recently described a multiparameter fluorescent aerolysin (FLAER)-based flow assay using CD45, CD33, and CD14 that accurately identified PNH monocyte and neutrophil clones in PNH, AA, and MDS. Here, we compared the efficiency of this WBC assay with a CD59-based assay on RBCs during a 3-year period. PNH clones were detected with the FLAER assay in 63 (11.8%) of 536 samples tested, whereas PNH RBCs were detected in only 33 (6.2%), and always with a smaller clone size. The FLAER assay on WBCs is a more sensitive and robust primary screening assay for detecting PNH clones in clinical samples.
authors.affiliation: Laboratory Medicine Program, Toronto General Hospital 11E-416, Toronto, Ontario M5G 2C4, Canada.
authors.forename: D Robert
authors.initials: DR
authors.lastname: Sutherland
authors.forename: Nancy
authors.initials: N
authors.lastname: Kuek
authors.forename: Juan
authors.initials: J
authors.lastname: Azcona-Olivera
authors.forename: Tanya
authors.initials: T
authors.lastname: Anderson
authors.forename: Erica
authors.initials: E
authors.lastname: Acton
authors.forename: David
authors.initials: D
authors.lastname: Barth
authors.forename: Michael
authors.initials: M
authors.lastname: Keeney
mesh_terms: D000741:Anemia, Aplastic / Q000276:immunology; D001427:Bacterial Toxins; D018957:CD59 Antigens / Q000032:analysis; D002999:Clone Cells; D004906:Erythrocyte Count / Q000379:methods; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D006457:Hemoglobinuria, Paroxysmal / Q000175:diagnosis* / Q000276:immunology; D006801:Humans; D007958:Leukocyte Count / Q000379:methods*; D009190:Myelodysplastic Syndromes / Q000276:immunology; D052899:Pore Forming Cytotoxic Proteins; D012680:Sensitivity and Specificity
chemical_list: D001427:Bacterial Toxins; D018957:CD59 Antigens; D052899:Pore Forming Cytotoxic Proteins; C032676:aerolysin; D016650:Fluorescein-5-isothiocyanate