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Protocol - Hemoglobin Characterization

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Protocol Name from Source:

This section will be completed when reviewed by an Expert Review Panel.

Availability:

Publicly available

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.

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, DNA sequencing analysis) that are appropriate to characterize hemoglobin variants. Once an assay is chosen for a particular study, the SRSP recommends that no changes in the protocol be made 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
  • E/A2
  • D-Punjab
  • G-Philadelphia
  • O-Arab

Personnel and Training Required

Phlebotomist

Equipment Needs

CLIA certified laboratory with the capability to perform the hemoglobin assay.

Requirements

Requirement CategoryRequired
Average time of greater than 15 minutes in an unaffected individualNo
Major equipmentNo
Specialized requirements for biospecimen collectionNo
Specialized trainingNo

Mode of Administration

Self-administered

Life Stage:

Infant, Toddler, Child, Adolescent, Adult, Senior, All Ages, Pregnancy

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 blood transfusions within the last 120 days.

The results of the Hemoglobin Characterization can be interpreted using the hemoglobinopathy case definition worksheets and classification tables from the Newborn Screening Technical Assistance and Evaluation Program (NewSTEPS, see [link[www.phenxtoolkit.org/toolkit_content/supplemental_info/scd_core_tier_1/measures/Sickle_Cell_Disease_Case_Definitions.doc|Sickle Cell Disease Case Definitions]]) funded by the Health Resources and Services Administration (HRSA). Investigators complete the case definition worksheets using available clinical data including: family history of hemoglobin variants, hemoglobin newborn screening results, laboratory record of complete blood count (CBC) with mean corpuscular volume (MCV), hemoglobin electrophoresis, isoelectric focusing, high performance liquid chromatography, and DNA genotype. Investigators then use the associated classification tables to interpret the results and make a diagnosis. The classification tables indicate the level of certainty (e.g., definite, probable, possible, or unlikely) of the diagnosis based on the type of clinical data available.

Research Domain Information

Release Date:

July 30, 2015

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.

Selection Rationale

The Laboratory Diagnosis of Haemoglobinopathies (1998) was compared to other protocols and selected by the Sickle Cell Disease Research and Scientific Panel (SRSP) because it includes advantages and disadvantages of the various techniques as well as the ability of the assays to separate the various hemoglobin variants.

Language

English

Standards

StandardNameIDSource
Common Data Elements (CDE)Hematology Hemoglobin F Measurement Assessment Finding Status Indicator4233132CDE Browser
Common Data Elements (CDE)Hematology Hemoglobin F Measurement Assessment Lab Unspecified Value3763197CDE Browser
Common Data Elements (CDE)Hematology Hemoglobin F Measurement Assessment Percentage Value3763134CDE Browser
Common Data Elements (CDE)Laboratory Procedure Hemoglobin Result Specified Lower Limit of Normal Value2003886CDE Browser
Common Data Elements (CDE)Laboratory Procedure Hemoglobin Result Specified Value2190CDE Browser
Common Data Elements (CDE)Laboratory Procedure Hemoglobin Result Unspecified Value2005035CDE Browser
Common Data Elements (CDE)Laboratory Procedure Hemoglobin Summary Result3113466CDE Browser

Process and Review

This section will be completed when reviewed by an Expert Review Panel.

Source

The Laboratory Diagnosis of Haemoglobinopathies. (1998). British Journal of Haematology, 101(4), 783-792. doi: 10.1046/j.1365-2141.1998.00809.x

General References

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:

830301

Variables:

Export Variables
Variable NameVariable IDVariable DescriptionVersiondbGaP Mapping
PX830301_SCD_Hemoglobin_Assay_ResultsPX830301060000Were the assay results recorded for the following hemoglobins? A, F, S, C, E/A2, D-Punjab, G-Philadelphia, O-Arab4N/A
PX830301_SCD_Hemoglobin_BloodDrawPX830301010000Was the blood drawn into an appropriate EDTA tube?4N/A
PX830301_SCD_Hemoglobin_Assay_Equipment_ManufacturerPX830301020000Who is the manufacturer of the equipment used?4N/A
PX830301_SCD_Hemoglobin_Assay_Equipment_MakePX830301030000What is the make of the equipment used?4N/A
PX830301_SCD_Hemoglobin_Assay_Equipment_CoefficientsPX830301040000What are the coefficients of variation for the assay?4N/A
PX830301_SCD_Hemoglobin_Assay_Equipment_RepeatabilityPX830301050000Describe the repeatability of the assay:4N/A