HB Electrophoresis Test: All you need to know about this test

Hemoglobin is present in the red corpuscles of our body and its function is to carry oxygen from the lungs to other organs and parts of the body. In a way, it acts like an oxygen carrier. There are two main types of haemoglobin found. Haemoglobin A which is the most commonly found and haemoglobin F which is the fetal haemoglobin that occurs in newborn babies. But after a while Hb F gets replaced with Hb A. if the haemoglobin levels in our blood is low then the person suffers from anemia.

There are some abnormal variety of haemoglobin seen like haemoglobin S which is found in case of sickle cell disease that leads to the formation of sickle shape of the red blood cells. These sickle shaped cells tend to get stuck in the blood vessels and cause pain and lack of oxygen leading to further complications. Haemoglobin C also is an abnormal type which is unable to deliver oxygen efficiently. Haemoglobin E also is seen in Asian race which lead to no or mild anemia.

Hemoglobin electrophoresis or hb electrophoresis is a test used to measure different types of haemoglobin be it normal or abnormal in our blood. In this an electric current passes through the blood sample. This differentiates the normal from abnormal haemoglobin.

In which conditions is the hb electrophoresis carried out?

Hb electrophoresis test can look up for abnormal variety of haemoglobin in the blood. So it is most useful in cases like anemia, sickle cell anemia, thalassemia, and other haemoglobin related disorders.

So who should take this test?

hemoglobin electrophoresis

If haemoglobin disorders struck you and you suffer from the below listed symptoms:

1. Tiredness

2. Yellow skin

3. Sickle cell disease

4. Growth related issues in children

Usually newborn babies are tested for any abnormality in their haemoglobin.

This is done to rule any abnormality if it exists at an early stage. Newborns undergo a lot of tests to check if any form of disease or abnormality exists in them.

You should also get yourself checked in case you have a family history of haemoglobin related disorders like thalassemia, sickle cell disease, specially if you are greek, belong to middle east, or Asian, African.

Hb electrophoresis in pregnancy can also be done to rule out any abnormal  haemoglobin present in the child.

hemoglobin electrophoresis procedure

Blood test Tube

The blood sample is collected by a lab technician or a healthcare professional from your vein in the arm. You will feel the prick of the needle, but this will be only for a few minutes and would go away quickly. For newborns the prick is made from the heal by rubbing alcohol on it. then after taking the blood that area is covered by a bandage.

Since electrophoresis is widely used for fractionation of various hemoglobin fractions, mainly because of differences in electric charge, the customary techniques of electrophoresis have been adapted for use with very small (pico- gram, 10-12 g) quantities in order to analyze hemoglobin mixtures. An electrophoresis method for picogram quantities has been developed by Edstrom for the analysis of RNA-DNA bases of single cells. The degree of separation of various haemoglobin fractions compares favourably with the separation obtained by polyacrylamide electrophoresis on a large scale.

The electrophoresis was conducted under direct microscopic observation in a special chamber. This chamber is made from a normal glass slide with glass tubes glued on the surface of it. a semicircular opening is present on the middle of the slide. Around one fourth volume of the tubes are filled with agarose solution in a buffer composed of glycine and HCL-tris. The agarose was connected through a semi-circular opening with a layer of agarose solution in the same buffer, which covers the slide surface between the tubes. After complete gelation of the agarose a slot was cut perpendicularly to the long axis of the agarose layer.

The agarose gels can be used as a buffer bridge to provide electrical connection with the extremities of acrylamide fibre. This fibre is represented by the track remaining in the middle of a collodion-coated coverslip after a thin stainless steel wire is pulled across a drop of acrylamide-erythrocyte suspension placed in the middle of the coverslip. Another drop of the same suspension is placed at the extremity of the track on the opposite edge of the coverslip. The acrylamide-erythrocyte suspension was prepared from a solution containing 20 percent acrylamide monomer (Eastman); 1 percent N, N’ methylenebis- acrylamide (Eastman), 10 percent glycerol. The erythrocytes are distributed at random in the fibre, and the number of cells per unit length of the fibre should be limited to avoid the overlapping of the haemoglobin fractions from various erythrocytes during the electrophoretic run.

What is the price of this test?

This test is very economical and can be done from any pathology lab from 500-1000 Rs.

Do you need to do any special preparations before the test?

No, no such special preparation or fasting is required for this test.

hemoglobin electrophoresis risks

hb electrophoresis test

you may feel a slight pinch in the area where the needle is put to withdraw blood and after that it might leave some bruise on that spot. This will heal in a few days. Newborns might also feel the pinch of the needle.

How to interpret the results?

The result will depict which type of abnormal haemoglobin exists.

Thalassemia is a condition where the overall production of haemoglobin is affected. Sickle cell disease is where there are s shape cells. A sickle cell gene is present. Haemoglobin C also is a form of mild anemia.