Genotype Test (Hemoglobin Electrophoresis Method)
The objective of the genotype test was to determine the genetic type of hemoglobin present in an individual, especially to identify conditions like sickle cell disease (SS, AS) or thalassemia.
2. Principle:
The test was based on the principle of electrophoresis, where different types of hemoglobin (e.g., HbA, HbS, HbC, HbF) migrated at different rates on an electrophoretic medium due to differences in their charge and structure.
3. Materials:
• EDTA blood sample
• Hemolysing reagent
• Electrophoresis apparatus (agarose or cellulose acetate gel)
• Power supply
• Buffer solution (alkaline pH)
• Staining dye
• Sample applicator
• Capillary tubes or pipettes
• Filter paper, gloves, and PPE
4. Procedure:
1. A venous blood sample was collected in an EDTA tube.
2. Red blood cells were lysed to release hemoglobin.
3. The hemolysate was applied to the electrophoresis gel strip.
4. Electrophoresis was carried out at alkaline pH using an electric current.
5. After separation, the bands were stained and interpreted by comparing with controls.
6. The migration pattern indicated the genotype (e.g., AA, AS, SS, AC)
.
5. Result:
• AA – Normal hemoglobin only
• AS – Sickle cell trait (carrier)
• SS – Sickle cell disease
• AC/SC – Other variant hemoglobins
6. Uses:
• It was used to screen for sickle cell disease, thalassemia, and other hemoglobinopathies.
• Also important for prenatal counseling, marriage counseling, and blood donor screening.
7. Consultation:
Patients with abnormal genotypes were referred to a hematologist or genetic counselor. Genetic counseling, preventive advice, or treatment plans were initiated based on the results.
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