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HelpOrder Code HBEL1 Hemoglobin Electrophoresis Evaluation, Blood
Ordering Guidance
Multiple hematology evaluations are available. For information on testing that is performed with each evaluation, see Benign Hematology Evaluation Comparison.
Necessary Information
At minimum, include recent transfusion information and most recent complete blood cell count results.
Metabolic Hematology Patient Information (T810) is strongly recommended. Testing may proceed without this information, however if the information requested is received, any pertinent reported clinical features and data will drive the focus of the evaluation and be considered in the interpretation.
The laboratory has extensive experience in hemoglobin variant identification and many cases can be confidently classified without molecular testing. However, molecular confirmation is always available, subject to sufficient sample quantity (eg, multiplex ligation-dependent probe amplification testing requires at least 2 mL of sample in addition to protein testing requirements). If no molecular testing or specific molecular tests are desired, utilize the appropriate check boxes on the form. If the form or other communication is not received, the reviewing hematopathologist will select appropriate tests to sufficiently explain the protein findings, which may or may not include molecular testing.
Specimen Required
Container/Tube:
Preferred: Lavender top (EDTA)
Acceptable: Yellow top (ACD solution B), green top (sodium heparin)
Specimen Volume: 10 mL
Collection Instructions: Send whole blood specimen in original tube. Do not aliquot.
Forms
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:
-Informed Consent for Genetic Testing (T576)
-Informed Consent for Genetic Testing-Spanish (T826)
2. Metabolic Hematology Patient Information (T810)
3. If not ordering electronically, complete, print, and send a Benign Hematology Test Request (T755) with the specimen
Useful For
Diagnosis and classification of hemoglobin disorders, including thalassemias and hemoglobin variants
Profile Information
| Test ID | Reporting Name | Available Separately | Always Performed |
|---|---|---|---|
| HBELI | Hb Electrophoresis Interpretation | No | Yes |
| HGBCE | Hb Variant, A2 and F Quantitation,B | Yes | Yes |
| HPLC | HPLC Hb Variant, B | No | Yes |
Reflex Tests
| Test ID | Reporting Name | Available Separately | Always Performed |
|---|---|---|---|
| HPFH | Hb F Distribution, B | No | No |
| MASS | Hb Variant by Mass Spec, B | No | No |
| SDEX | Sickle Solubility, B | Yes | No |
| IEF | Isoelectric Focusing, B | No | No |
| UNHB | Hb Stability, B | No | No |
| ATHAL | Alpha-Globin Gene Analysis | Yes | No |
| WASQR | Alpha Globin Gene Sequencing, B | Yes, (Order WASEQ) | No |
| WBSQR | Beta Globin Gene Sequencing, B | Yes, (Order WBSEQ) | No |
| WBDDR | Beta Globin Cluster Locus Del/Dup,B | Yes, (Order WBDD) | No |
| WGSQR | Gamma Globin Full Gene Sequencing | Yes, (Order WGSEQ) | No |
| HBEL0 | Hb Electrophoresis Summary Interp | No | No |
Testing Algorithm
This evaluation will always include hemoglobin (Hb) A2 and HbF and hemoglobin electrophoresis utilizing capillary electrophoresis and cation exchange high-performance liquid chromatography methods.
Reflex testing, performed at additional charge, may include any or all of the following to identify rare hemoglobin variants present: sickle solubility (hemoglobin S screen); hemoglobin heat and isopropanol stability studies (unstable hemoglobin); isoelectric focusing, intact globin chain mass spectrometry (hemoglobin variant by mass spectrometry); HbF distribution by flow cytometry; DNA Sanger sequencing assays for: 1) beta-chain variants and the most common beta thalassemias (beta-globin gene sequencing), 2) alpha-chain variants and less common nondeletional alpha thalassemias (alpha-globin gene sequencing), or 3) gamma-chain variants and nondeletional hereditary persistence of fetal hemoglobin (HPFH) (gamma-globin full gene sequencing); multiplex ligation-dependent probe amplification assays for: 1) large deletional alpha thalassemias and alpha-gene duplications (alpha-globin gene analysis), or 2) beta-globin gene cluster locus large deletions and duplications, including large deletional HPFH, delta-beta thalassemia, gamma-delta-beta thalassemia, epsilon-gamma-delta-beta thalassemia and large deletional beta or delta thalassemia (beta-globin cluster locus deletion/duplication).
If test results in the profile are abnormal, results may be reviewed by a hematopathology consultant, and a summary interpretation provided.
One or more of the following molecular tests may be reflexed:
-ATHAL / Alpha-Globin Gene Analysis, Varies
-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, Varies
For cases with molecular testing added, a preliminary interpretation will be reported that discusses the protein test results. After all test results are finalized, an additional consultative interpretation that summarizes all testing and incorporates subsequent genetic results will be provided.
Special Instructions
Method Name
HBELI, HBEL0: Medical Interpretation
HGBCE: Capillary Electrophoresis
HPLC: Cation Exchange/High-Performance Liquid Chromatography (HPLC)
IEF: Isoelectric Focusing
MASS: Mass Spectrometry (MS)
HPFH: Flow Cytometry
UNHB: Isopropanol and Heat Stability
Reporting Name
Hb Electrophoresis EvaluationSpecimen Type
Whole Blood EDTASpecimen Minimum Volume
1 mL (this volume will limit reflex testing possibilities)
3 mL if multiplex ligation-dependent probe amplification is needed
Specimen Stability Information
| Specimen Type | Temperature | Time | Special Container |
|---|---|---|---|
| Whole Blood EDTA | Refrigerated | 7 days |
Reject Due To
All specimens will be evaluated at Mayo Clinic Laboratories for test suitability.Reference Values
Hemoglobin Electrophoresis Interpretation
Definitive results and an interpretative report will be provided.
Hemoglobin Variant, A2 and F Quantitation
HEMOGLOBIN A
0-30 days: 5.9-77.2%
1-2 months: 7.9-92.4%
3-5 months: 54.7-97.1%
6-8 months: 80.0-98.0%
9-12 months: 86.2-98.0%
13-17 months: 88.8-98.0%
18-23 months: 90.4-98.0%
≥24 months: 95.8-98.0%
HEMOGLOBIN A2
0-30 days: 0.0-2.1%
1-2 months: 0.0-2.6%
3-5 months: 1.3-3.1%
≥6 months: 2.0-3.3%
HEMOGLOBIN F
0-30 days: 22.8-92.0%
1-2 months: 7.6-89.8%
3-5 months: 1.6-42.2%
6-8 months: 0.0-16.7%
9-12 months: 0.0-10.5%
13-17 months: 0.0-7.9%
18-23 months: 0.0-6.3%
≥24 months: 0.0-0.9%
VARIANT 1
0.0
VARIANT 2
0.0
VARIANT 3
0.0
Method Description
Hemoglobin Electrophoresis:
The CAPILLARYS System is an automated system that uses capillary electrophoresis to separate charged molecules by their electrophoretic mobility in an alkaline buffer. Separation occurs according to the electrolyte pH and electro-osmotic flow. A sample dilution with hemolyzing solution is injected by aspiration. A high-voltage protein separation occurs with direct detection of the hemoglobin-protein fractions at 415 nm, which is specific to hemoglobin. The resulting electrophoregram peaks are evaluated for pattern abnormalities and are quantified as a percentage of the total hemoglobin present. Examples of position of commonly found hemoglobin fractions are, from cathode to anode: HbA2', C, A2/O-Arab, E, S, D, G-Philadelphia, F, A, Hope, Barts, J, N-Baltimore and H.(Louahabi A, Philippe M, Lali S, Wallemacq P, Maisin D: Evaluation of a new Sebia kit for analysis of hemoglobin fractions and variants on the Capillarys system. Clin Chem Lab Med. 2006;44[3]:340-345; instruction manual: CAPILLARYS Hemoglobin(E) using the CAPILLARYS 2 flex-piercing instrument. Sebia; 06/2014)
High-Performance Liquid Chromatography:
Hemolysate of whole blood is injected into an analysis stream passing through a cation exchange column using high-performance liquid chromatography (HPLC). A preprogrammed gradient controls the elution buffer mixture that also passes through the analytical cartridge. The ionic strength of the elution buffer is raised by increasing the percentage of a second buffer. As the ionic strength of the buffer increases the more strongly retained hemoglobins elute from the cartridge. Absorbance changes are detected by a dual-wavelength filter photometer. Changes in absorbance are displayed as a chromatogram of absorbance versus time.(Huismann TH, Scroeder WA, Brodie AN, Mayson SM, Jakway J: Microchromotography of hemoglobins. III. A simplified procedure for the determination of hemoglobin A2. J Lab Clin Med. 1975;86:700-702; Ou CN, Buffone GJ, Reimer GL, Alpert AJ: High-performance liquid chromatography of human hemoglobins on a new cation exchanger. J Chromatogr. 1983;266:197-205; Szuberski J, Oliveira JL, Hoyer JD: A comprehensive analysis of hemoglobin variants by high-performance liquid chromatography [HPLC]. Int J Lab Hematol. 2012 Dec; 34(6):594-604; instruction manual: Bio-Rad Variant II Beta-thalassemia Short Program Instructions for Use, L70203705. Bio-Rad Laboratories, Inc; 11/2011)
Performing Laboratory
Mayo Clinic Laboratories in Rochester
CPT Code Information
83020
83021
82664 (if appropriate)
83068 (if appropriate)
83789 (if appropriate)
88184 (if appropriate)
83020-26 (if appropriate)
Day(s) Performed
Monday through Thursday