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Solubility Test For Sickle Cell

Sickle cell anemia is caused by a mutation in the hemoglobin gene that results in abnormal hemoglobin S production. Red blood cells with hemoglobin S change shape in low oxygen conditions from round discs to rigid sickle shapes. This limits blood flow and oxygen transport, causing anemia and potentially life-threatening conditions. Sickle cell disease can be screened for via blood tests that induce sickling through reducing conditions, identifying turbidity caused by hemoglobin S crystal formation.

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Christian Oranza
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574 views14 pages

Solubility Test For Sickle Cell

Sickle cell anemia is caused by a mutation in the hemoglobin gene that results in abnormal hemoglobin S production. Red blood cells with hemoglobin S change shape in low oxygen conditions from round discs to rigid sickle shapes. This limits blood flow and oxygen transport, causing anemia and potentially life-threatening conditions. Sickle cell disease can be screened for via blood tests that induce sickling through reducing conditions, identifying turbidity caused by hemoglobin S crystal formation.

Uploaded by

Christian Oranza
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd
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Practical

Clinical Hematology
:What is Sickle cell
Sickle cell anemia is an inherited disorder that leads to
the production of an abnormal hemoglobin variant,
hemoglobin S (HbS)

Hemoglobin S production arises from an altered


(mutated) “S” gene. Differ on Hb A by one amino acid
substitution. .
Amino acid substitution in HbS
RBCs have Hbs
In the RBC, variant Hbs can form polymers in low
oxygen conditions, changing the shape of the RBC
from a round disc to a characteristic crescent (sickle)
shape.
This altered shape limits the RBC’s ability to flow
smoothly throughout the body, limits the
hemoglobin’s ability to transport oxygen, and
decreases the RBCs lifespan from 120 days to about 10-
20 days.
The affected person can become anemic because the
body cannot produce RBCs as fast as they are
destroyed.
 Also, sickled blood cells can become trapped in blood
vessels reducing or blocking blood flow. This can
damage organs, muscles, and bones and may lead to
life-threatening conditions.
Sickle cell test:
A sickle cell test is a blood test done to screen for
sickle cell trait or sickle cell disease.
There are three method
1. A saline citrate with paraffin oil.
2. Sodium Metabisulfite Method.
3. Solubility Test.
General Principle
We will make the conditions at which oxygen tension
decline to induced the sickling process of Hbs in RBCs.
A saline citrate with paraffin oil
Principle:
a saline citrate suspension of blood is allowed to stand
in a test tube under a layer of paraffin oil until sickling
takes place.’
In employing any of the common diagnostic tests for
sickling it is desirable to obtain blood which has a low.
fraction of oxyhemoglobin.
Sodium Metabisulfite Method
Principle
When a drop of blood is sealed between a
cover slip and a slide, the decline in oxygen
tension due to oxidative processes in the blood
cells leads to sickling.
In this method added with blood drop a
chemical reducing agents. Such as sodium
metabisulfite. This rapidly reduces
oxyhemoglobin to reduced hemoglobin, then
this will be accelerate sickling .
Solubility test
Principle
Erythrocytes are lysed by saponin and the released
hemoglobin is reduced by sodium hydrosulfite in a
concentrated phosphate buffer.
Under these conditions, reduced HbS is characterized
by its very low solubility and the formation of crystals.
Procedure
Blood is drawn into a tube

Sodium dithionite or a similar chemical like (saponin ,


.etc.) is then added to the blood

Positive: The plasma will become cloudy because the


.hemoglobin S is actually forming small crystals
Principle
The presences of HbS or HbC are indicated by the
turbid solutions. The normal HbA under these same
conditions results in a clear non-turbid solutions.

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