1. SICKLE CELL ANEMIA
Nada Mohamed Ahmed ,
MD, MT (ASCP)i

2. HEMOLYTIC ANEMIA Hereditary
Intracorpuscular
WITHIN THE RED CELL
HEMOLYTIC ANEMIA
Extracorpuscular
OUTSIDE THE RED CELL
Hereditary
Acquired
AUTO-IMMUNE
1. Warm Ab
2. Cold Ab
3. Transfusion reactions
4. Drug associated
NON-IMMUNE
1. Hypersplenism
2. Infections (Malaria),,
3-mechanical trauma to RBCs
4. Liver dz (Spur cell)
1. Membrane defects
- Hereditary spherocytosis
- Hereditary elliptocytosis
- Hereditary pyropoikilocytosis
- Hereditary stomatocytosis
2. Enzyme defects
-G6PD
3. -Hemoglobin defects
-.Hemoglbinopathies(sickle cell disorders)
- Hb SS, CC, SC & S-B-
- Thalassemias
Membrane defects
PNH

3. OBJECTIVES
Definition Causes and Genetics design Classification Pathogenesis Lab diagnosis

4. What is Sickle cell Disease
Sickle Cell disease: is a genetic disorder that affects erythrocytes (RBC) causing them to become sickle or crescent shaped.
The effects of this condition due to an abnormality of the hemoglobin molecules found in erythrocytes.

5. The origin of Sickle Cell anemia
The change in cell structure arises from a change in
the structure of hemoglobin.
A single change in an amino acid causes hemoglobin
to aggregate.

6. Genetics design It’s autosomal recessive blood disease.
You inherit it from your parents.
*The gene defect is a known mutation of a single nucleotide.
*The person that receives the defective gene from both his parents will develop Sickle-cell disease.
An autosomal recessive disorder means two copies of an abnormal gene must be present in order for the disease or trait to develop.
*The person who receives only one defective gene from either one of his parents will develop Sickle-cell trait.

7. Causes Sickle Cell
Is caused by a point mutation in the β-globin chain of haemoglobin, causing the amino acid glutamic acid to be replaced with the c amino acid valine at the sixth position.

8. Types of Hemoglobin
Normal
Hb A
Hb A2
Hb F
Abnormal (350)
Hb S
Hb C
Hb E

9. Haemoglobin A two alpha two beta A2 two delta F two gamma
Normally, humans have
Of these, Haemoglobin A makes up around 96-97% of the normal haemoglobin in humans.
Haemoglobin A
two alpha
two beta A2
two delta F
two gamma

10. Classification Sickle Cell Disease
Hemoglobin SS Disease
Hemoglobin SC Disease
Sickle Cell Trait

11. Common types of Sickle Cell Disorders
Classification
Common types of Sickle Cell Disorders
Type of anaemia
Hemoglobin variation
comment
Sickle Cell Anemia
Sickle haemoglobin (HbS) + Sickle haemoglobin
Most Severe – No HbA
Hemoglobin S-C disease
Sickle haemoglobin (HbS) + (HbC)
Mild form of Sickle Cell Disorder
Sickle Cell Trait
Sickle haemoglobin (S) + Normal haemoglobin (A)

12. Sickle Cell Anemia vs. Sickle Cell Trait
People who have sickle cell anemia are born with it; means inherited, lifelong condition. They inherit two copies of sickle cell gene, one from each parent. Sickle cell trait is different from sickle cell anemia . People with sickle cell trait don’t have the condition, but they have one of the genes that cause the condition. People with sickle cell anemia and sickle cell trait can pass the gene on when they have children.
Sickle cell anemia is an inherited, lifelong condition. People who have sickle cell anemia are born with it. They inherit two copies of the sickle cell gene, one from each parent. People who inherit a sickle cell gene from one parent and a normal gene from the other parent have a condition called sickle cell trait.
Sickle cell trait is different from sickle cell anemia. People with sickle cell trait don’t have the condition, but they have one of the genes that cause the condition. Like people with sickle cell anemia, people with sickle cell trait can pass the gene on when they have children. To learn more about sickle cell trait, see the section on causes of sickle cell anemia.

13. Sickle Cell Trait
Sickle haemoglobin (S) + Normal haemoglobin (A) in RBC
Adequate amount of normal Hb (A) in red blood cells
RBC remain flexible
Carrier
Do Not have the symptoms of the sickle cell disorders,
with exceptions:
Pain when Less Oxygen than usual
Minute kidney problems

14. Mechanism of Pathogenesis
When sickle hemoglobin (HbS) gives up its oxygen to the tissues, HbS sticks together
Forms long rods form inside RBC
RBC become rigid, inflexible, and sickle-shaped
Unable to squeeze through small blood vessels, instead blocks small blood vessels
Less oxygen to tissues of body
RBCs containing HbS have a shorter lifespan
Normally 20 days
Chronic state of anaemia

18. Red Blood Cells from Sickle Cell Anemia
Deoxygenation of SS erythrocytes leads to intracellular hemoglobin polymerization, loss of deformability and changes in cell morphology.
OXY-STATE
DEOXY-STATE

19. Diagnosis Complete blood count Sickle test solubility tests
hemoglobin electrophoresis test
DNA Analysis
Screening test for newborns

20. Laboratory investigations
Complete blood count
Level of Hb -: 6–8 g/dL
(Normal range-: Male= g/dl Female= g/dl)
High reticulocyte count (10–20%).
Blood film
The blood film is microcytic and hypocromic
Sickled cell anaemia
Normal

22. This test is simple and quick, used to identify the presence of HbS.
Sickling Test
Method:
1) A sample of venous blood or capillary blood may be collected for this test.
2) Mixing blood with the reducing agent, sodium metabisulphite, will induce sickling in susceptible cells.
3) the results can be viewed under a microscope after 20 minutes.
This test is simple and quick, used to identify the presence of HbS.
Negative Test
HbA
Positive Test
HbS
Normal RBC
Sickled RBC

23. Sickle Solubility Test (SST)
A rapid and inexpensive technique used to screen for the presence of sickling hemoglobins, can be used at home.
A positive result must be confirmed by another method (HPLC or electrophoresis) to confirm the presence of Hb S and to distinguish Hb AS (carrier state) from Hb SS (sickle cell disease).
Disadvantage: Other insoluble hemoglobins, such as Hb C-Harlem, will also give a positive result.
Method
1) Erythrocytes are lysed by saponin.
2) The released hemoglobin is reduced by sodium hydrosulfite in a phosphate buffer.
3) Reduced HbS is characterized by its turbidity ( insolubility).

24. Sickle Solubility Test (SST
The presence of HbA under these same conditions results in a clear red solution. 
The resulting tactoids of HbS causes the solution to remain turbid. 

25. Hemoglobin Electrophoresis test
* Haemoglobin electrophoresis will differentiate between homozygous and heterozygous conditions.
* Hemoglobin types have different electrical charges and move at different speeds.
*HbAS: Has both HbA and HbS.
Shows 2 bands
*HbSS: Is less negative by 2 compared to HbA .
Migrates slower than HbA

26. DNA analysis
 This test is used to investigate alterations and mutations in the genes that produce hemoglobin components.
It may be performed to determine whether someone has one or two copies of the Hb S mutation or has two different gene mutations.
Genetic testing is most often used for prenatal testing:
amniocentesis “14 to 16 weeks”.

27. Screening 16 and 18 weeks of the pregnancy
Prenatal Testing
Amniocentesis
16 and 18 weeks of the pregnancy
small risk of causing a miscarriage (1 in 100)

28. Signs and Symptoms of Sickle Cell Anemia
Jaundice
Pain Episodes
Anemia
Infections
Ulcers on the Legs
Eye Problems

29. What Are the Signs and Symptoms of Sickle Cell Anemia?
The signs and symptoms of sickle cell anemia vary. Some people have mild symptoms. Others have very severe symptoms and often are hospitalized for treatment. Sickle cell anemia is present at birth, but many infants don’t show any signs until after 4 months of age.

30. Signs and Symptoms Related to Anemia
The most common symptom of anemia is fatigue (feeling tired or weak). Other signs and symptoms of anemia include:
Shortness of breath
Dizziness
Headache
Coldness in the hands and feet
Pale skin
Chest pain

31. What Are the Signs and Symptoms of Sickle Cell Anemia?
Signs and Symptoms Related to Pain Sudden pain throughout the body is a common symptom of sickle cell anemia. This pain is called a "sickle cell crisis." Sickle cell crises often affect the bones, lungs, abdomen, and joints. A sickle cell crisis occurs when sickled red blood cells form clumps in the bloodstream. (Other cells also may play a role in this clumping process.) These clumps of cells block blood flow through the small blood vessels in the limbs and organs. This can cause pain and organ damage.