Protocol - Hemoglobin Characterization
Description
This protocol provides basic instructions for drawing and storing blood and performing the bioassay for hemoglobin characterization. Because there are many assays that may be required to characterize hemoglobin abnormalities, the protocol also provides basic guidelines to increase comparability among different studies.
Specific Instructions
The Sickle Cell Disease Research and Scientific Panel (SRSP) recommends that investigators also capture Complete Blood Count and Reticulocyte Count.
The SRSP recommends that results from Hemoglobin Characterization be interpreted in the context of age-appropriate normal values and history of blood transfusions within the last 120 days.
The results of Hemoglobin Characterization can be interpreted using the algorithms in the Association of Public Health Laboratories and Center for Disease Control and Prevention document: " Hemoglobinopathies: Current Practices for Screening, Confirmation and Follow-up". Characterization is based on the results of hemoglobin electrophoresis, isoelectric focusing, high performance liquid chromatography, capillary zone electrophoresis, immunoassays, and DNA genotype. More definitive diagnosis may require complete blood count (CBC) with mean corpuscular volume (MCV), reticulocyte count, and family studies.
Published guidelines on hemoglobinopathy interpretation all focus on newborn screening results, however, the Sickle Cell Disease Research and Scientific Panel (SRSP) suggests that they can be used in adult populations by modifying the algorithms by removing fetal hemoglobin if it is not the dominant hemoglobin (e.g., "FS" becomes "S", "FSC" becomes "SC", "FSA" becomes "SA", "FAS" becomes "AS" etc.).
Availability
Protocol
Hemoglobin Characterization
Blood Draw
Blood should be drawn into an appropriate EDTA tube.
Laboratory Assay for Hemoglobin Characterization
The Sickle Cell Disease Scientific and Research Panel (SRSP) notes that there are numerous different assays and instruments (e.g., hemoglobin electrophoresis, isoelectric focusing, high performance liquid chromatography, capillary zone electrophoresis, DNA sequencing analysis) that are appropriate to characterize hemoglobin variants. Once an assay is chosen for a particular study, the SRSP recommends that the same protocol be used over the course of the study. To aid comparability, the SRSP recommends that the investigator record the make and manufacturer of equipment used and the repeatability and coefficients of variation for the assay.
The SRSP notes that investigators should record the assay results for the following hemoglobins (the exact list of hemoglobins that can be distinguished will depend on the assay being used). The hemoglobins are listed in order of the amount present with the most prevalent coming first.
- A
- F
- S
- C
- A2
- E
- D
- G
- O
Personnel and Training Required
Phlebotomist
Equipment Needs
CLIA certified laboratory with the capability to perform the hemoglobin assay.
Requirements
| Requirement Category | Required |
|---|---|
| Major equipment | No |
| Specialized training | No |
| Specialized requirements for biospecimen collection | No |
| Average time of greater than 15 minutes in an unaffected individual | No |
Mode of Administration
Bioassay
Lifestage
Infant, Toddler, Child, Adolescent, Adult, Senior, Pregnancy
Participants
All ages
Selection Rationale
The Association of Public Health Laboratories and Center for Disease Control and Prevention document: "Hemoglobinopathies: Current Practices for Screening, Confirmation and Follow-up" was compared to other protocols and selected by the Sickle Cell Disease Research and Scientific Panel (SRSP) because it discusses advantages and disadvantages of the various techniques as well as the ability of the assays to separate the various hemoglobin variants.
Language
English
Standards
| Standard | Name | ID | Source |
|---|
Derived Variables
None
Process and Review
The PhenX Sickle Cell Disease Research and Scientific Panel (SRSP) reviewed this protocol in May 2022. Guidance from the SRSP includes:
- Updated protocol
Protocol Name from Source
Association of Public Health Laboratories (APHL)
Source
Association of Public Health Laboratories (APHL). Hemoglobinopathies: Current Practices for Screening, Confirmation and Follow-up. https://www.aphl.org/aboutAPHL/publications/Documents/NBS_HemoglobinopathyTesting_122015.pdf. Accessed April 26, 2022
General References
CLSI. Newborn Screening for Hemoglobinopathies. 1st ed. CLSI guideline NBS08. Wayne, PA: Clinical and Laboratory Standards Institute; 2019.
The Laboratory Diagnosis of Haemoglobinopathies. (1998). British Journal of Haematology, 101(4), 783-792. doi: 10.1046/j.1365-2141.1998.00809.x
Kutlar, F. (2007). Diagnostic approach to hemoglobinopathies. Hemoglobin, 31(2), 243-250.
Kutlar, A., & Huisman, T. (1996). Detection of hemoglobinopathies. In F. Hommes (Ed.), Techniques in Diagnostic Human Biochemical Genetics: A Laboratory Manual (pp. 519-560). New York, New York: J. Wiley & Sons.
Protocol ID
830302
Variables
Export Variables| Variable Name | Variable ID | Variable Description | dbGaP Mapping | |
|---|---|---|---|---|
| PX830302_Hemoglobin_Characterization_Assay_Equipment_Coefficients | ||||
| PX830302040000 | What are the coefficients of variation for more | N/A | ||
| PX830302_Hemoglobin_Characterization_Assay_Equipment_Make | ||||
| PX830302030000 | What is the make of the equipment used? | N/A | ||
| PX830302_Hemoglobin_Characterization_Assay_Equipment_Manufacturer | ||||
| PX830302020000 | Who is the manufacturer of the equipment used? | N/A | ||
| PX830302_Hemoglobin_Characterization_Assay_Equipment_Repeatability | ||||
| PX830302050000 | Describe the repeatability of the assay: | N/A | ||
| PX830302_Hemoglobin_Characterization_Assay_Results | ||||
| PX830302060000 | Were the assay results recorded for the more | N/A | ||
| PX830302_Hemoglobin_Characterization_Assay_Results_List | ||||
| PX830302070000 | What were the assay results for the more | N/A | ||
| PX830302_Hemoglobin_Characterization_Blood_Draw | ||||
| PX830302010000 | Was the blood drawn into an appropriate EDTA tube? | N/A | ||
Measure Name
Hemoglobin Characterization
Release Date
October 30, 2023
Definition
A bioassay for hemoglobin classification.
Purpose
This protocol can be used to identify and characterize the different variants in structure and synthesis of hemoglobin that cause sickle cell disease.
Keywords
sickle cell disease, SCD, SC disease, anemia, thalassemia, hemoglobin, hemoglobinopathy, high-performance liquid chromatography, HPLC, isoelectric focusing, IEF, immunoassay
Measure Protocols
| Protocol ID | Protocol Name |
|---|---|
| 830302 | Hemoglobin Characterization |
Publications
There are no publications listed for this protocol.