Ordering Recommendation

Preferred test for initial diagnosis of paroxysmal nocturnal hemoglobinuria (PNH) and quantification of PNH clones.

Mnemonic

PNH PAN

Methodology

Quantitative Flow Cytometry

Performed

Sun-Sat

Reported

1-3 days

New York DOH Approval Status

This test is New York DOH approved.

Specimen Required

Patient Preparation
Collect

Lavender (EDTA), pink (K2EDTA), or green (sodium or lithium heparin).

Specimen Preparation

Transport 4 mL whole blood. (Min: 4 mL)

Storage/Transport Temperature

Refrigerated.

Unacceptable Conditions

Clotted or hemolyzed specimens.

Remarks

Specimens must be analyzed within stability times provided.

Stability

Ambient: 24 hours; Refrigerated: 72 hours; Frozen: Unacceptable
New York State Clients: Ambient: 24 hours; Refrigerated: 48 hours. Frozen: Unacceptable

Reference Interval

By report

Interpretive Data

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 2018 (Cytometry B Clin. Cytom. 2018; 94B:49). The test includes high-sensitivity WBC and RBC analysis with a lower limit of quantification of 0.02 percent for PNH RBCs and PMNs (based on 250,000 cells analyzed) and 0.5 percent for PNH monocytes (based on 10,000 cells analyzed). The lower limit of detection for PNH RBCs and PMNs is 0.008 percent and for PNH monocytes 0.2 percent. For severely pancytopenic patients, the WBC assay sensitivity will be much lower.

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).

Patient Retesting Recommendations: The frequency of testing is dictated by clinical and hematologic parameters; repeat testing is indicated upon any significant change in clinical or laboratory parameters and is suggested at least annually for routine monitoring. In the setting of aplastic anemia, international guidelines recommend screening for PNH at diagnosis, and every 3 to 6 months initially, reducing the frequency of testing if the proportion of GPI-deficient cells has remained stable over an initial two-year period (Int J Lab Hematol 2019;41 Suppl 1:73-81).

This test was developed and its performance characteristics determined by ARUP Laboratories. It has not been cleared or approved by the U.S. Food and Drug Administration. This test was performed in a CLIA-certified laboratory and is intended for clinical purposes.

Compliance Category

Laboratory Developed Test (LDT)

Note

If >1% PNH RBCs are detected, then PNH RBC TYPE reflex will be added at no additional charge.

Hotline History

N/A

CPT Codes

86356 x7

Components

Component Test Code* Component Chart Name LOINC
2004367 Total (II and III) CD59-deficient RBC 33662-8
2005004 FLAER and CD157-deficient monocytes 77948-8
2005005 FLAER and CD157-deficient neutrophils 77948-8
3005033 RBC PNH Phenotype 93479-4
3005034 Neutrophil PNH Phenotype 93479-4
3005035 Monocyte PNH Phenotype 93479-4
* Component test codes cannot be used to order tests. The information provided here is not sufficient for interface builds; for a complete test mix, please click the sidebar link to access the Interface Map.

Aliases

  • CD157, FLAER, CD59, Glycophorin A
  • PI-Linked, eculizumab monitoring
Paroxysmal Nocturnal Hemoglobinuria (PNH), High Sensitivity, RBC and WBC