Paroxysmal Nocturnal Hemoglobinuria (PNH) Screen
Label Mnemonic: LAB8255
Order form: Epic Beaker AP Order Label
Flow Cytometry Service
6240 RCP
Flow Cytometry Resident: 467-6008
Flow Cytometry Lab: 467-6009 or 467-6010
Tube Station #260
Peripheral Blood
Collection Medium:
Yellow top tube 8.5 mL (ACD solution A)
Adult or pediatric: 10 mL; yellow top tube (ACD-A)
Delivery Instructions:
Deliver to laboratory immediately after collection. Send to tube station #260 or deliver to Specimen Control, 6240 RCP.
Testing Schedule:
0800-1630 Monday through Friday. For additional services, contact Clinical Pathology Resident on-call at pager #3404.
Turn Around Time:
2 days
Reference Range:
An interpretative report will be provided by the pathologist.
Ham's acid hemolysin and sucrose lysis tests have been replaced 
by flow cytometric testing for glycosylphosphatidyl inositol (GPI)-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). 

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. 
Flow Cytometry
CPT Code:
Technical:     88184 x1 and 88185 x6
Professional:  88187
See Additional Information:
Specimens Requiring Immediate Delivery