Test Catalog

Test ID: RBCME    
Red Blood Cell Membrane Evaluation, Blood

Useful For Suggests clinical disorders or settings where the test may be helpful

Investigation of suspected red cell membrane disorders such as hereditary spherocytosis or hereditary pyropoikilocytosis

Testing Algorithm Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

Osmotic fragility and eosin-5-maleimide (EMA) binding (Band3) flow cytometry testing will always be performed. A normal shipping control is necessary to exclude false-positive results due to preanalytical artifact. Testing will be canceled if no shipping control is received or if the shipping control is abnormal. A consultative interpretation will be provided.


See Benign Hematology Evaluation Comparison in Special Instructions.

Clinical Information Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test

The functional red cell membrane is composed of a cholesterol and phospholipid bilayer anchored by integral proteins to an elastic cytoskeletal network. These interactions form the shape, deformability, and proper ion balance of the cell. Abnormalities in these moieties result in red blood cell membrane disorders.


Hereditary spherocytosis (HS) is a common membrane disorder that can be present in many ethnic groups. Its prevalence has been estimated at approximately 1 in 3,000 persons of Northern European ancestry. It is usually associated with visible spherocytes on the peripheral blood smear and can be associated with variable clinical features of hemolysis ranging from completely compensated to mild to severe.


Hereditary elliptocytosis (HE) is another fairly common and clinically variable disorder that can range from normal RBC indices in the large majority of cases to rare patients with moderate to severe anemia.


Common hereditary elliptocytosis (CHE) is characterized by the presence of elliptocytes on the peripheral blood smear. Mutations associated with HE have been reported in widely variable ethnicities with greater prevalence in populations overlapping the malaria belt.


Hereditary pyropoikilocytosis (HPP) is best classified as a severe form of hereditary elliptocytosis. It is uncommon and presents in early childhood as a severe hemolytic anemia. These disorders are associated with marked poikilocytosis on the peripheral blood smear.(1,2) Red cell membrane disorders can result from abnormalities involving several red cell membrane proteins, such as band 3, alpha and beta spectrin, protein 4.1, protein 4.2, glycophorin C, and ankyrin.


Most often, red cell membrane disorders are diagnosed in childhood, adolescence, or early adult life. The diagnosis of HS is usually made by a combination of patient and family history, laboratory evidence of hemolysis, and review of a peripheral blood smear. The osmotic fragility (OF) test is usually markedly abnormal in HS cases. However, factors such as age, iron status, and medications can affect the OF test. The OF test is nonspecific and can be increased in acquired disorders such as autoimmune hemolytic anemia. Coombs testing should be negative prior to ordering this test.


The addition of eosin-5-maleimide (EMA) binding (Band3) flow cytometry to this profile increases specificity if a typical moderately decreased pattern is present. Hereditary pyropoikilocytosis can have normal or only mildly increased OF results and often displays a very dim and sometimes dual peak pattern with EMA-binding testing. Common hereditary elliptocytosis cases are not discriminated from normal patients in either OF and EMA binding (band3) testing and this profile does not add confirmatory information for HE.

Reference Values Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.

> or =12 months:

0.50 g/dL NaCl (unincubated): 3-53% hemolysis

0.60 g/dL NaCl (incubated): 14-74% hemolysis

0.65 g/dL NaCl (incubated): 4-40% hemolysis

0.75 g/dL NaCl (incubated): 1-11% hemolysis 

An interpretive report will be provided.


Reference values have not been established for patients who are <12 months of age.

Interpretation Provides information to assist in interpretation of the test results

An interpretive report will be provided.

Cautions Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances

This test is not useful for hereditary elliptocytosis.

Interfering factors:

-Recent transfusion

-Oral contraceptives

-H2 blockers

Infrequently, other hemolytic disorders may also be associated with positive osmotic fragility results, as in patients with congenital nonspherocytic hemolytic anemia due to glucose-6-phosphate dehydrogenase or pyruvate kinase deficiency. Eosin-5-maleimide (EMA) binding (band 3) results can be masked by reticulocytosis. Results must be interpreted within the clinical, familial, and peripheral blood smear findings.


Patients with an immunohemolytic anemia, or who have recently received a blood transfusion may also have increased RBC lysis.


The shipping control specimen is required to adequately interpret these cases, as temperature extremes can increase fragility of the specimen. 


Resulting Cautions:

Osmotic fragility results will be reported if the shipping control is normal.


If the shipping control is abnormal and the osmotic fragility results are within normal range, the results will be reported; however, a comment will be added to the report indicating that the shipping control was not entirely satisfactory.


The test will be cancelled if the patient specimen and shipping control are both abnormal.

Clinical Reference Recommendations for in-depth reading of a clinical nature

1. 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

2. Lux SE, IV: 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

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

4. 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

Special Instructions Library of PDFs including pertinent information and forms related to the test