Preferred test for initial diagnosis of paroxysmal nocturnal hemoglobinuria (PNH) and quantification of PNH clones.
Quantitative Flow Cytometry
Lavender (EDTA), pink (K2EDTA), or green (sodium or lithium heparin).
Transport 4 mL whole blood. (Min: 4 mL)
Bone marrow. Clotted or hemolyzed specimens.
Specimens must be analyzed within stability times provided.
Ambient: 24 hours; Refrigerated: 72 hours; Frozen: Unacceptable
New York State Clients: Ambient: 24 hours; Refrigerated: 48 hours. Frozen: Unacceptable
This test is preferred for the initial diagnosis of PNH, and was developed according to published guidelines (Cytometry B Clin. Cytom. 2010; 78:211) and as updated in 2014 (Cytometry B Clin. Cytom. 2014; 86:44). The test includes high-sensitivity WBC and RBC analysis with lower limits of detection of 0.005 percent for RBCs, 0.005 percent for PMNs, and 0.020 percent for monocytes.
WBC analysis is the most accurate measurement of the PNH clone size. FLAER and CD157 are used as GPI-linked markers; CD15 (PMNs) and CD64 (monocytes) are used as lineage-specific markers. RBC analysis quantifies Type II and Type III RBC clones when the percentage of PNH RBCs is greater than 1 percent. Glycophorin A (CD235a) is used to gate the RBC population, and CD59 is the GPI-linked antigen. Recent RBC transfusions may decrease the percentage of PNH cells measured in RBCs (Cytometry 2000; 42:223). The presence of a subclinical PNH population in myelodysplastic bone marrow disorders, such as aplastic anemia or refractory anemia, may correlate with a positive immunotherapeutic response (Blood 2006; 107, 1308-1314).
Laboratory Developed Test (LDT)
|Component Test Code*||Component Chart Name||LOINC|
|2004367||% PNH RBC||33662-8|
|2005004||% PNH Monocytes||60554-3|
|2005005||% PNH PMN||53831-4|
- CD15, CD33, CD14, CD24, FLAER, CD59, Glycophorin A
- PI-Linked Antigens, RBCs, Monocytes and Granulocytes