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Test ID: HAEVP Hemolytic Anemia Evaluation

Reporting Name

Hemolytic Anemia Evaluation

Useful For

Evaluation of lifelong or inherited hemolytic anemias, including red cell membrane disorders, unstable or abnormal hemoglobin variants, and red cell enzyme disorders


This evaluation is not suitable for acquired causes of hemolysis.

Clinical Information

Hemolytic anemia (HA) is characterized by increased red cell destruction and a decreased red cell life span. Patients usually have decreased hemoglobin concentration, hematocrit, and red blood cell count, but some can have compensated disorders, and symptoms such as reticulocytosis, pigmented gallstones, and decreased haptoglobin are factors that raise clinical suspicion. Blood smear abnormalities may include spherocytes, schistocytes, stomatocytes, polychromasia, basophilic stippling, and target cells. Osmotic fragility can be increased due to the presence of spherocytes. These are all nonspecific features that can be present in both hereditary and acquired hemolytic disorders.


Inherited hemolytic disorders may include red cell membrane disorders, red cell enzyme defects, or abnormalities in the hemoglobin molecule in the red cell. This panel assesses for possible causes of congenital/hereditary causes of hemolytic anemia and does not evaluate for acquired causes. Therefore, the anemia should be lifelong or familial in nature. Examples of acquired HA (which should be excluded prior to ordering this panel) include: autoimmune HA (direct Coombs-positive HA, Coombs-negative autoimmune HA), cold agglutinin disease, paroxysmal nocturnal hemoglobinuria, paroxysmal cold hemoglobinuria, mechanical hemolysis (aortic stenosis or prosthetic heart valves), disseminated intravascular coagulation/thrombotic microangiopathy, and drug-induced HA.


This consultation evaluates for a hereditary cause of increased red cell destruction and includes testing for red cell membrane disorders, such as hereditary spherocytosis and hereditary pyropoikilocytosis, hemoglobinopathies, and red cell enzyme abnormalities.


This panel is of limited use in patients with a history of recent transfusion and should be ordered as remote a date from transfusion as possible in those patients who are chronically transfused.


An interpretive report will be provided.

Testing Algorithm

This is a consultative evaluation in which the case will be evaluated and appropriate tests performed, at an additional charge, and the results interpreted. If a peripheral blood smear is provided, the morphologic features will be incorporated into the interpretation. If a Hemolytic Anemia Patient Information sheet (T705) is received with the sample, the reported clinical features or clinical impression will be incorporated into the interpretation.


The most common RBC enzymes (G6PD,pyruvate kinase, glucose phosphate isomerase, and hexokinase) will always be performed. If these are normal, the second-tier enzymes will be performed (provided sufficient sample volume is available). If second-tier enzymes are desired, even if the first-tier testing is abnormal, fill out the Hemolytic Anemia Patient Information sheet (T705) and indicate this desire. Cation exchange HPLC, capillary electrophoresis, and hemoglobin stability studies will always be performed. Reflex testing required to identify a hemoglobin abnormality can be added as the case requires. Osmotic fragility and eosin-5-maleimide (EMA) binding (band 3) flow cytometry will be performed on all cases. A normal shipping control for osmotic fragility (OF) is necessary to exclude false-positive results due to preanalytical artifact.


OF and EMA binding testing will be canceled if no shipping control is received or if the shipping control is abnormal.


HAEVA / Hemolytic Anemia Summary Interpretation, an additional consultative interpretation that summarizes all testing, will be provided after test completion to incorporate subsequent results into an overall evaluation if 1 or more of the following molecular tests are reflexed on the Hemolytic Anemia Evaluation:

-ATHAL / Alpha-Globin Gene Analysis

-WASQR / Alpha-Globin Gene Sequencing, Blood

-WBSQR / Beta-Globin Gene Sequencing, Blood

-WBDDR / Beta-Globin Cluster Locus Deletion/Duplication, Blood

-WGSQR / Gamma-Globin Full Gene Sequencing


Note: RBCE / Reflexed RBC Enzymes, Blood (second-tier enzymes) includes: adenylate kinase, phosphofructokinase, phosphoglycerate kinase, triosephosphate isomerase, and pyrimidine 5' nucleotidase.


See Benign Hematology Evaluation Comparison in Special Instructions.

Profile Information

Test ID Reporting Name Available Separately Always Performed
HAEV Hemolytic Anemia Interpretation No Yes
A2F Hemoglobin A2 and F No Yes
HBEL Hemoglobin Electrophoresis, B No Yes
UNHB Unstable Hemoglobin, B No Yes
FRAGO Osmotic Fragility Yes, (Order FRAG) Yes
SCTRL Shipping Control Vial No Yes
G6PD G-6-PD, QN, RBC Yes Yes
PK Pyruvate Kinase, RBC Yes Yes
GPI Glucose Phosphate Isomerase, B Yes Yes
HEXK Hexokinase, B No Yes
PBSM Morphology Review No Yes

Reflex Tests

Test ID Reporting Name Available Separately Always Performed
GLTI Glutathione, B No No
SDEX Hemoglobin S, Scrn, B Yes No
IEF IEF Confirms No No
MASS Hb Variant by Mass Spec, B No No
RBCE Reflexed RBC Enzymes No No
HPFH Hemoglobin F, Red Cell Distrib, B No No
ATHAL Alpha-Globin Gene Analysis Yes No
WASQR Alpha Globin Gene Sequencing, B Yes, (Order WASEQ) No
WBDDR Beta Globin Cluster Locus Del/Dup,B Yes, (Order WBDD) No
HAEVA Hemolytic Anemia Summary Interp No No
WBSQR Beta Globin Gene Sequencing, B No No
WGSQR Gamma Globin Full Gene Sequencing No No

Analytic Time

3-25 days if structural and/or molecular studies are required (not reported Saturday or Sunday)

Day(s) and Time(s) Performed

Monday through Saturday

Clinical Reference

1. Steiner LA, Gallagher PG: Erythrocyte disorders in the perinatal period. Semin Perinatol 2007 Aug;31(4):254-261. PMID: 17825683

2. Beutler E: Glucose-6-phosphate dehydrogenase deficiency and other enzyme abnormalities. In Hematology. Fifth edition. Edited by E Beutler, MA Lichtman, BS Coller, TJ Kipps. New York, McGraw-Hill Book Company, 1995, pp 564-581

3. Hoyer JD, Hoffman DR: The thalassemia and hemoglobinopathy syndromes. In Clinical Laboratory Medicine. Second edition. Edited by KD McMlatchey. Philadelphia, Lippincott, Williams and Wilkins, 2002, pp 866-895

4. King MJ, Garcon L, Hoyer JD, et al: International Council for Standardization in Haematology. ICSH guidelines for the laboratory diagnosis of nonimmune hereditary red cell membrane disorders. Int J Lab Hematol. 2015 Jun;37(3):304-325. PMID: 25790109

5. Lux SE: Anatomy of the red cell membrane skeleton: unanswered questions. Blood 2016 Jan 14;127(2):187-199 doi: 10.1182/blood-2014-12-512772. PMID: 26537302

6. Gallagher PG: Abnormalities of the erythrocyte membrane. Pediatr Clin North Am 2013 Dec;60(6):1349-1362. PMID: 24237975

7. Bianchi P, Fermo E, Vercellati C, et al: Diagnostic power of laboratory tests for hereditary spherocytosis: a comparison study in 150 patients grouped according to molecular and clinical characteristics. Haematologica 2012 Apr;97(4):516-523. PMID: 22058213

8. Cappellini MD, Fiorelli G: Glucose-6-phosphate dehydrogenase deficiency. Lancet 2008;371:64-74

9. Hereditary hemolytic anemias due to red blood cell enzyme disorders. Edited by B Glader. Philadelphia: Wolters Kluwer/Lippincott, Williams and Wilkins; 2014, pp 728

Method Name

HAEV: Consultative Interpretation

A2F: Cation Exchange/High-Performance Liquid Chromatography (HPLC)

HBEL: Capillary Electrophoresis

UNHB: Isopropanol and Heat Stability

FRAGO: Osmotic Lysis

G6PD, PK, GPI, HEXK, RBCE, GLTI: Kinetic Spectrophotometry (KS)

PBSM: Consultant Review

BND3 (eosin-5-maleimide (EMA) binding): Flow Cytometry

MASS: Mass Spectrometry (MS)

IEF: Electrophoresis

HPFH: Flow Cytometry

HAEVA: Consultative Interpretation

Specimen Type

Whole Blood ACD-B
Whole Blood EDTA
Whole Blood Slide

Advisory Information

Preliminary screening tests, such as complete blood count with peripheral smear and direct Coombs test with a negative result, should be run before ordering this evaluation.


Cold agglutinin disorders and autoimmune disorders should be excluded prior to testing. This evaluation is not suitable for acquired causes of hemolysis.

Shipping Instructions

Specimens must arrive within 72 hours of draw.

Necessary Information

Include recent transfusion information.


Include most recent CBC results.

Specimen Required

Two whole blood EDTA specimens, 2 whole blood ACD specimens, an EDTA control specimen, and 2 well-made peripheral blood smears (Wright stained or fixed in absolute methanol) are required for testing.



Specimen Type: Blood

Container/Tube: Lavender top (EDTA) and yellow top (ACD)

Specimen Volume:

EDTA: Two 4-mL vials

ACD: Two 6-mL vials

Collection Instructions:

1. Immediately refrigerate specimens after draw.

2. Send specimens in original tubes. Do not aliquot.

3. Prepare smears.

4. Rubber band patient specimen and control vial together.



Specimen Type: Slides

Container/Tube: Blood smears

Specimen Volume: 2 well-made peripheral blood smears

Collection Instructions: Collect 2 well-made peripheral blood smears (Wright stained or fixed in absolute methanol).


Shipping Normal Control:

Specimen Type: Whole blood

Container/Tube: Lavender top (EDTA)

Specimen Volume: 4 mL

Collection Instructions:

1. Draw a control specimen from a normal (healthy), unrelated, nonsmoking person at the same time as the patient.

2. Label clearly on outermost label normal control.

3. Immediately refrigerate specimen after draw.

4. Send specimen in original tube. Do not aliquot.

5. Rubber band patient specimen and control vial together.

Specimen Minimum Volume

EDTA Blood: 3 mL
ACD Blood: 5 mL

Specimen Stability Information

Specimen Type Temperature Time Special Container
Control Refrigerated 72 hours PURPLE OR PINK TOP/EDTA
Whole Blood ACD-B Refrigerated 72 hours
Whole Blood EDTA Refrigerated 72 hours
Whole Blood Slide Refrigerated CARTRIDGE

Reference Values

Definitive results and an interpretive report will be provided.

Test Classification

See Individual Test IDs

CPT Code Information

Hemolytic Anemia Evaluation

82657-Hexokinase, B


83020-Hemoglobin electrophoresis

83021-Hemoglobin A2 and F

83068-Hemoglobin stability

84087-Glucose phosphate isomerase

84220-Pyruvate kinase

85060-Morphology review

85557-Osmotic fragility


Band 3 Fluorescence Staining, RBC



Reflexed RBC Enzymes

83915 (if appropriate)


Glutathione, Blood

82978 (if appropriate)


Hemoglobin Variant by Mass Spectrometry (MS), Blood

83789 (if appropriate)


IEF Confirms

82664 (if appropriate)


Hemoglobin F, Red Cell Distribution, Blood

88184 (if appropriate)

LOINC Code Information

Test ID Test Order Name Order LOINC Value
HAEVP Hemolytic Anemia Evaluation In Process


Result ID Test Result Name Result LOINC Value
HEXK_ Hexokinase, B 49216-5
PK_ Pyruvate Kinase, RBC 32552-2
9095 Hemoglobin, Unstable, B 4639-1
9064 Osmotic Fragility, RBC 34964-7
G6PD_ G-6-PD, QN, RBC 32546-4
GPI_ Glucose Phosphate Isomerase, B 44050-3
2380 Hemoglobin A 20572-4
13082 Morphology Review 59466-3
2381 Hemoglobin A2 42245-1
SCTRL Shipping Control Vial 40431-9
9992 Hemolytic Anemia Interpretation 59466-3
37437 Reviewed By 19139-5
2382 Hemoglobin F 42246-9
2383 Variant 32017-6
3306 Osmotic Fragility, 0.50 g/dL NaCl 23915-2
3307 Osmotic Fragility, 0.60 g/dL NaCl 23918-6
29224 Variant 2 32017-6
29225 Variant 3 32017-6
3308 Osmotic Fragility, 0.65 g/dL NaCl 23920-2
3309 Osmotic Fragility, 0.75 g/dL NaCl 23921-0
2101 Interpretation 78748-1
3310 Osmotic Fragility Comment 59466-3


1. New York Clients-Informed consent is required. Document on the request form or electronic order that a copy is on file. The following documents are available in Special Instructions:

-Informed Consent for Genetic Testing (T576)

-Informed Consent for Genetic Testing-Spanish (T826)

2. Metabolic Hematology Patient Information (T810) in Special Instructions. Please fill out for a more complete evaluation by the signing Hematopathologist.

3. If not ordering electronically, complete, print, and send a Benign Hematology Test Request (T755) with the specimen.

Additional Tests

Test ID Reporting Name Available Separately Always Performed
BND3 Band 3 Fluorescence Staining, RBC No Yes
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