|Order form:||Epic Beaker AP Order Label|
Flow Cytometry Resident: 467-6008
Flow Cytometry Lab: 467-6009 or 467-6010
Tube Station #260
|Yellow top tube 8.5 mL (ACD solution A)|
Ham's acid hemolysin and sucrose lysis tests have been replaced by flow cytometric testing for glycosylphosphatidyl inositol (GPI)- anchored proteins CD14 and CD24, and aerolysin binding. These proteins are not expressed on PNH white blood cells and their lack of expression is determined by flow cytometric assay. The channel-forming toxin, aerolysin, and its inactive precursors, proaerolysin, bind selectively with a high affinity to the GPI anchor itself. The lack of GPI anchor on blood cell surface will decrease the ability of fluorescently labeled protein aerolysin (FLAER) to bind to nucleated blood cells in patients with PNH. Determination of CD14 and CD24 must be performed on fresh whole blood. Both monocytes and granulocytes are analyzed for CD14/CD24 expression and aerolysin bindings. Granulocytes are the most sensitive population in which to detect GPI-anchored protein deficiency. Two additional markers are performed for gating purposes, CD45 (leukocyte common antigen) and CD15 (myeloid antigen). REFS: 1)Richards, S et al. Application of Flow Cytometry to the Diagnosis of Paroxysmal Nocturnal Hemoglobinuria. Cytometry 2000; 42:223-233. 2)Dunn, D, et al. Paroxysmal Nocturnal Hemoglobinuria in Patients with Bone Marrow Failure Syndromes. Ann Int Med 1999; 131:401-408. 3) Brodsky RA, et al. Improved detection and characterization of paroxysmal nocturnal hemoglobinuria using fluorescent aerolysin. Am J Clin Pathol 2000; 114:459-66.
Technical: 88184 x1 and 88185 x6 Professional: 88187
Specimens Requiring Immediate Delivery