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Test Code PLINK Paroxysmal Nocturnal Hemoglobinuria, PI-Linked Antigen, Blood

Reporting Name

PNH, PI-Linked AG, B

Useful For

Screening for and confirming the diagnosis of paroxysmal nocturnal hemoglobinuria (PNH)


Monitoring patients with PNH

Performing Laboratory

Mayo Clinic Laboratories in Rochester

Specimen Type

Whole blood

Specimen Required

Specimen must arrive within 72 hours of draw.



Preferred: Yellow top (ACD)

Acceptable: Lavender top (EDTA)

Specimen Volume: 2.6 mL

Collection Instructions: Send specimen in original tube. Do not transfer blood to other containers.

Specimen Minimum Volume

1 mL

Specimen Stability Information

Specimen Type Temperature Time Special Container
Whole blood Ambient (preferred) 72 hours
  Refrigerated  72 hours

Reference Values

An interpretive report will be provided. 



PNH RBC-Partial Antigen loss: 0.00-0.99%

PNH RBC-Complete Antigen loss: 0.00-0.01%

PNH Granulocytes: 0.00-0.01%

PNH Monocytes: 0.00-0.05%

Day(s) and Time(s) Performed

Specimens are processed Monday through Sunday.

Results reported Monday through Friday.

Test Classification

This test was developed using an analyte specific reagent. Its performance characteristics were determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the U.S. Food and Drug Administration.

CPT Code Information

88184-Flow cytometry, RBC x 1

88184-Flow cytometry, WBC x 1

88185-Flow cytometry, additional marker (each), RBC x 1

88185-Flow cytometry, additional marker (each), WBC x 6

88188-Flow Cytometry Interpretation, 9-15 Markers x 1

LOINC Code Information

Test ID Test Order Name Order LOINC Value
PLINK PNH, PI-Linked AG, B 90735-2


Result ID Test Result Name Result LOINC Value
CK079 Interpretation 90739-4
CK080 PNH RBC-Partial Ag Loss 33662-8
CK081 PNH RBC-Complete Ag Loss 90738-6
CK082 PNH Granulocytes 90737-8
CK083 PNH Monocytes 90736-0

Clinical Reference

1. Miyata T, Yamada N, Iida Y, et al: Abnormalities of PIG-A transcripts in granulocytes from patients with paroxysmal nocturnal hemoglobinuria. N Engl J Med. 1994;330:249-255

2. Brodsky RA: Advances in the diagnosis and therapy of paroxysmal nocturnal hemoglobinuria. Blood Rev. 2008 Mar;22(2):65-74

3. Richards SJ, Barnett D: The role of flow cytometry in the diagnosis of paroxysmal nocturnal hemoglobinuria in the clinical laboratory. Clin Lab Med. 2007 Sep;27(3):577-590

4. Parker C, Omine M, Richards S, et al: Diagnosis and management of paroxysmal nocturnal hemoglobinuria. Blood. 2005 Dec 1;106(12):3699-3709

5. Richards SJ, Hill A, Hillman P: Recent advances in the diagnosis, monitoring and management of patients with paroxysmal nocturnal hemoglobinuria. Cytometry B Clin Cytom. 2007 Sep;72(5):291-298

6. Shichishima T, Terasawa T, Hashimoto C, et al: Discordant and heterogeneous expression of GPI-anchored membrane proteins on leukemic cells in a patient with paroxysmal nocturnal hemoglobinuria. Blood. 1993;81:1855-1862

7. Brodsky RA, Mukhina GL, Li S, et al: Improved detection and characterization of paroxysmal nocturnal hemoglobinuria using fluorescent aerolysin. Am J Clin Pathol. 2000;114:459-466

8. Rosse WF: Phosphatidylinositol-linked proteins and paroxysmal nocturnal hemoglobinuria. Blood. 1990;75:1595-1601

9. Borowitz MJ, Craig FE, DiGiuseppe JA, et al: Guidelines for the diagnosis and monitoring of paroxysmal nocturnal hemoglobinuria and related disorders by flow cytometry. Cytometry B Clin Cytom. 2010 78(4):211-230

10. van der Schoot CE, Huizinga TW, van't Veer-Korthof ET, Wijmans R, Pinkster J, von dem Born AEe: Deficiency of glycosyl-phosphatidylinositol-linked membrane glycoproteins of leukocytes in paroxysmal nocturnal hemoglobinuria, description of a new diagnostic cytofluorometric assay. Blood. 1990;76:1853-1859

Method Description

Flow cytometric immunophenotyping of peripheral blood (WBC and RBC) is performed using the following antibodies;


RBC: CD235a, CD59 

WBC: CD14, CD15, CD16, CD24, CD33, CD45, and FLAER 


This assay evaluates the presence or absence of glycosylphosphatidylinositol (GPI)-linked proteins using monoclonal antibodies directed against CD235, CD33, and CD15 to isolate different cell lineages. GPI-linked proteins that are checked within different lineages include CD14 for monocytes, CD's 16 and 24 for granulocytes, and CD59 for RBCs. Fluorescent aerolysin, a fluorescently labeled inactive variant of the protein aerolysin binds selectively to GPI anchors and is also evaluated for presence or absence of expression on WBCs. In addition, this test will detect a partial loss of CD59 on RBCs (type II RBCs).


Individuals without paroxysmal nocturnal hemoglobinuria have normal expression of all GPI-linked antigens on peripheral blood and leukocytes and erythrocytes.(Richards SJ, Hill A, Hillman P: Recent advances in the diagnosis, monitoring and management of patients with paroxysmal nocturnal hemoglobinuria. Cytometry B Clin Cytom 2007 Sep;72[5]:291-298)

Analytic Time

1 day

Reject Due To

Gross hemolysis Reject

Method Name


Additional Tests

Test ID Reporting Name Available Separately Always Performed
FCIMS Flow Cytometry Interp, 9-15 Markers No, (Bill Only) Yes

Secondary ID