Ann Lab Med.  2015 Jan;35(1):35-40. 10.3343/alm.2015.35.1.35.

A Novel Marker for Screening Paroxysmal Nocturnal Hemoglobinuria Using Routine Complete Blood Count and Cell Population Data

• Kahng J
• Kim Y
• Kim JO
• Koh K
• Lee JW
• Han K

• KMID: 2363145
• DOI: http://doi.org/10.3343/alm.2015.35.1.35

Abstract

BACKGROUND
Final diagnosis of paroxysmal nocturnal hemoglobinuria (PNH) may take years demanding a quick diagnosis measure. We used the facts that PNH cells are damaged in acid, and reagents for measuring reticulocytes in Coulter DxH800 (Beckman Coulter, USA) are weakly acidic and hypotonic, to create a new PNH screening marker.
METHODS
We analyzed 979 complete blood counts (CBC) data from 963 patients including 57 data from 44 PNH patients. Standard criteria for PNH assay for population selection were followed: flow cytometry for CD55 and CD59 on red blood cells (RBCs) to a detection level of 1%; and fluorescent aerolysin, CD24 and CD15 in granulocytes to 0.1%. Twenty-four PNH minor clone-positive samples (minor-PNH+) were taken, in which the clone population was <5% of RBCs and/or granulocytes. Excluding PNH and minor-PNH+ patients, the population was divided into anemia, malignancy, infection, and normal groups. Parameters exhibiting a distinct demarcation between PNH and non-PNH groups were identified, and each parameter cutoff value was sought that includes the maximum [minimum] number of PNH [non-PNH] patients.
RESULTS
Cutoff values for 5 selected CBC parameters (MRV, RDWR, MSCV, MN-AL2-NRET, and IRF) were determined. Positive rates were: PNH (86.0%), minor-PNH+ (33.3%), others (5.0%), anemia (13.4%), malignancy (5.3%), infection (3.7%), normal (0.0%); within anemia group, aplastic anemia (40.0%), immune hemolytic anemia (11.1%), iron deficiency anemia (1.6%). Sensitivity (86.0%), specificity (95.0%), PPV (52.1%), and NPV (99.1%) were achieved in PNH screening.
CONCLUSION
A new PNH screening marker is proposed with 95% specificity and 86% sensitivity. The flag identifies PNH patients, reducing time to final diagnosis by flow cytometry.

Keyword

Paroxysmal nocturnal hemoglobinuria; Screening marker; Complete blood count; Reticulocyte-related parameters

MeSH Terms

Antigens, CD15/metabolism
Antigens, CD24/metabolism
Antigens, CD55/metabolism
Antigens, CD59/metabolism
Biomarkers/metabolism
Blood Cell Count
Erythrocytes/cytology/metabolism
Flow Cytometry
Granulocytes/cytology/metabolism
Hemoglobinuria, Paroxysmal/*diagnosis/metabolism
Humans
Sensitivity and Specificity
Antigens, CD15
Antigens, CD24
Antigens, CD55
Antigens, CD59
Biomarkers

Figure

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  Fig. 1 Examples of flow cytometric analysis for red blood cells in a paroxysmal nocturnal hemoglobinuria (PNH) patient with a clone size of 93.7% (A) and another PNH patient with a clone size of 4.9% (B).

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