title: Diagnosis of paroxysmal nocturnal hemoglobinuria in peripheral blood and bone marrow with six-color flow cytometry.
abstract: OBJECTIVE
  Identification of paroxysmal nocturnal hemoglobinuria (PNH) by detecting a glycophosphatidylinositol-anchored defect by flow cytometry is presently the standard method of choice for diagnosing PNH. However, the selection of suitable markers will be critical and significantly affect the determination and quantification of PNH clones in various cell lineages.
  
  
  METHODS
  In this study, we investigated the performance of various immunophenotypic markers including CD59, GPHA (a clustered antigen, CD235a), CD33, CD15 and fluorescent aerolysin (FLAER) combined with CD16, CD24 and CD14 in a PNH panel using six-color flow cytometry.
  
  
  RESULTS
  The results strongly indicate that these markers can collectively and effectively identify and quantify PNH clones in erythrocyte, granulocyte and monocyte populations derived from peripheral blood and bone marrow (BM). A sensitivity threshold as low as 0.01% in identifying PNH clones in erythrocyte and granulocyte populations from peripheral blood is achieved by this panel in a series dilution assay. In addition, a direct side-by-side comparison between BM and peripheral blood from the same patients suggests that the FLAER PNH test is capable of identifying to PNH clones in BM specimens.
  
  
  CONCLUSIONS
  The data support the premise that a six-color flow cytometry PNH panel using the combination of CD59, CD235a, CD33, CD15, FLAER, CD16, CD24 and CD14 can enhance and improve the current methods used in diagnosis and management of PNH by specifically identifying PNH clones in the erythrocyte, granulocyte and monocyte population.
authors: Esoterix Genetic Laboratories LLC, 521 W 57th Street, New York, NY 10019, USA. yangh3@labcorp.com
authors: Hai-Su
authors: HS
authors: Yang
authors: Min
authors: M
authors: Yang
authors: Xiaoyu
authors: X
authors: Li
authors: Sorina
authors: S
authors: Tugulea
authors: Henry
authors: H
authors: Dong
mesh_terms: D000328:Adult; D000368:Aged; D001427:Bacterial Toxins / Q000737:chemistry; D001772:Blood Cell Count; D001853:Bone Marrow / Q000378:metabolism* / Q000473:pathology; D051927:CD24 Antigen / Q000378:metabolism; D018957:CD59 Antigens / Q000378:metabolism; D019070:Cell Lineage; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D058851:GPI-Linked Proteins / Q000378:metabolism; D006098:Granulocytes / Q000166:cytology / Q000378:metabolism; D006457:Hemoglobinuria, Paroxysmal / Q000097:blood / Q000175:diagnosis*; D006801:Humans; D016130:Immunophenotyping; D016256:Lewis X Antigen / Q000378:metabolism; D018950:Lipopolysaccharide Receptors / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000166:cytology / Q000378:metabolism; D052899:Pore Forming Cytotoxic Proteins / Q000737:chemistry; D017452:Receptors, IgG / Q000378:metabolism; D063268:Sialic Acid Binding Ig-like Lectin 3 / Q000378:metabolism
chemical_list: D001427:Bacterial Toxins; D051927:CD24 Antigen; D018957:CD59 Antigens; D058851:GPI-Linked Proteins; D016256:Lewis X Antigen; D018950:Lipopolysaccharide Receptors; D052899:Pore Forming Cytotoxic Proteins; D017452:Receptors, IgG; D063268:Sialic Acid Binding Ig-like Lectin 3; C032676:aerolysin