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

Profile Information
A profile is a group of laboratory tests that are ordered and performed together under a single Mayo Test ID. Profile information lists the test performed, inclusive of the test fee, when a profile is ordered and includes reporting names and individual availability.

Test Id Reporting Name Available Separately Always Performed
FRAGO Osmotic Fragility Yes, (Order FRAG) Yes
SCTRL Shipping Control Vial No Yes
HSEV Spherocytosis Interpretation No Yes
BND3 Band 3 Fluorescence Staining, RBC No Yes
SMPB Peripheral Blood Smear Review No Yes

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.

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

Method Name
A short description of the method used to perform the test

HSEV: Consultative Interpretation

FRAGO: Osmotic Lysis

BND3: Flow Cytometry

SMPB: Consultant Review

NY State Available
Indicates the status of NY State approval and if the test is orderable for NY State clients.

Yes

Reporting Name
Lists a shorter or abbreviated version of the Published Name for a test

RBC Membrane Evaluation, B

Aliases
Lists additional common names for a test, as an aid in searching

Band 3

EMA Binding

Osmotic Fragility

Eosin-5-maleimide

Eosin

RBC membrane

Hereditary spherocytosis

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.

Specimen Type
Describes the specimen type validated for testing

Control

Whole Blood EDTA

Whole Blood Slide

Shipping Instructions

Specimens must arrive within 72 hours of draw.

Necessary Information

Include recent transfusion information.

 

Include most recent CBC results.

Specimen Required
Defines the optimal specimen required to perform the test and the preferred volume to complete testing

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

 

Patient:

Specimen Type: Blood

Container/Tube: Lavender top (EDTA)

Specimen Volume: 4 mL

Collection Instructions:

1. Immediately refrigerate specimen after draw. Refrigerate at 0 to 4 degrees C. Do not freeze. Freezing causes sample lysis, and tests will not be performed on hemolyzed specimens.

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

3. Rubber band patient specimen and control vial together. Control must accompany the patient sample at all times to ensure the reliability of testing results.

4. Be sure specimen and control are stored and transported together at refrigerate temperature, carefully following proper handling and shipping instructions.

 

Patient:

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).

 

Normal Shipping 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. Refrigerate at 0 to 4 degrees C. Do not freeze. Freezing causes sample lysis, and tests will not be performed on hemolyzed specimens.

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

5. Rubber band patient specimen and control vial together. Control must accompany the patient sample at all times to ensure the reliability of testing results.

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

Forms

1. Metabolic Hematology Patient Information (T810) in Special Instruction

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

Specimen Minimum Volume
Defines the amount of sample necessary to provide a clinically relevant result as determined by the Testing Laboratory

2 mL

Reject Due To
Identifies specimen types and conditions that may cause the specimen to be rejected

Gross hemolysis Reject
Other Clotted Frozen

Specimen Stability Information
Provides a description of the temperatures required to transport a specimen to the performing laboratory, alternate acceptable temperatures are also included

Specimen Type Temperature Time Special Container
Control Refrigerated (preferred) 72 hours PURPLE OR PINK TOP/EDTA
Whole Blood EDTA Refrigerated (preferred) 72 hours
Whole Blood Slide Refrigerated (preferred) CARTRIDGE

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

Method Description
Describes how the test is performed and provides a method-specific reference

Osmotic Fragility:

Specimens for erythrocyte osmotic fragility tests are anticoagulated with EDTA. Osmotic lysis is performed using sodium chloride (NaCl) solution, 0.50 g/dL. An incubated fragility test is performed following 24-hour incubation at 37 degrees C at the following NaCl concentrations: 0.60, 0.65, and 0.75 g/dL. Results are reported and interpreted.(Larson CJ, Scheidt R, Fairbanks VF: The osmotic fragility test for hereditary spherocytosis: use of EDTA-anticoagulated blood stored at 4 degrees C for up to 96 hours. Am Soc Clin Pathol Meeting Abstract, 1988; Larson CJ, Scheidt R, Fairbanks VF: The osmotic fragility test for hereditary spherocytosis: objective criteria for test interpretation. Am Soc Clin Pathol Meeting Abstract, 1988)

 

Band 3:

Eosin-5-maleimide (EMA) is a fluorescent dye that binds to Lys-430 of the extracellular loop of the band 3 protein. Using a 1-color flow cytometry method (number of events plotted against fluorescence), the fluorescent intensity of EMA-stained RBC, is assessed and compared to normal-value patients.(King MJ, Behrens J, Rogers C, et al: Rapid flow cytometric test for the diagnosis of membrane cytoskeletal associated hemolytic anemia. Br J Haematol 2000;111:924-933)

 

Peripheral Blood Smear Review:
A hematopathologist who is an expert in these disorders evaluates the slides and an interpretive report is issued.

PDF Report
Indicates whether the report includes an additional document with charts, images or other enriched information

No

Day(s) Performed
Outlines the days the test is performed. This field reflects the day that the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time before the test is performed. Some tests are listed as continuously performed, which means that assays are performed multiple times during the day.

Monday through Saturday

Report Available
The interval of time (receipt of sample at Mayo Clinic Laboratories to results available) taking into account standard setup days and weekends. The first day is the time that it typically takes for a result to be available. The last day is the time it might take, accounting for any necessary repeated testing.

4 to 7 days

Specimen Retention Time
Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded

7 days

Performing Laboratory Location
Indicates the location of the laboratory that performs the test

Rochester

Fees
Several factors determine the fee charged to perform a test. Contact your U.S. or International Regional Manager for information about establishing a fee schedule or to learn more about resources to optimize test selection.

  • Authorized users can sign in to Test Prices for detailed fee information.
  • Clients without access to Test Prices can contact Customer Service 24 hours a day, seven days a week.
  • Prospective clients should contact their Regional Manager. For assistance, contact Customer Service.

Test Classification
Provides information regarding the medical device classification for laboratory test kits and reagents. Tests may be classified as cleared or approved by the US Food and Drug Administration (FDA) and used per manufacturer instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR) product.

This test was developed, and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the US Food and Drug Administration.

CPT Code Information
Provides guidance in determining the appropriate Current Procedural Terminology (CPT) code(s) information for each test or profile. The listed CPT codes reflect Mayo Clinic Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.

CPT codes are provided by the performing laboratory.

85557-Osmotic fragility

88184-Flow cytometry; first cell surface, cytoplasmic or nuclear marker x 1

85060-Morphology review

LOINC® Information
Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the order and results codes of this test. LOINC values are provided by the performing laboratory.

Test Id Test Order Name Order LOINC Value
RBCME RBC Membrane Evaluation, B 98905-3
Result Id Test Result Name Result LOINC Value
Applies only to results expressed in units of measure originally reported by the performing laboratory. These values do not apply to results that are converted to other units of measure.
83141 Band 3 Fluorescence Staining, RBC 98906-1
9064 Osmotic Fragility, RBC 34964-7
3306 Osmotic Fragility, 0.50 g/dL NaCl 23915-2
3307 Osmotic Fragility, 0.60 g/dL NaCl 23918-6
3308 Osmotic Fragility, 0.65 g/dL NaCl 23920-2
3309 Osmotic Fragility, 0.75 g/dL NaCl 23921-0
3310 Osmotic Fragility Comment 59466-3
SCTRL Shipping Control Vial 40431-9
13065 Spherocytosis Interpretation 50595-8
37436 Reviewed By 18771-6
37406 Peripheral Blood Smear Review 59465-5

Test Setup Resources

Setup Files
Test setup information contains test file definition details to support order and result interfacing between Mayo Clinic Laboratories and your Laboratory Information System.

Excel | Pdf

Sample Reports
Normal and Abnormal sample reports are provided as references for report appearance.

Normal Reports | Abnormal Reports

SI Sample Reports
International System (SI) of Unit reports are provided for a limited number of tests. These reports are intended for international account use and are only available through MayoLINK accounts that have been defined to receive them.

SI Normal Reports | SI Abnormal Reports