A SEMINAR PRESENTATION

ON
SICKLE CELL ANEMIA AND IT’S MEDICAL LABORATORY DIAGNOSIS

PRESENTED BY:
ADARERHI OGHENERHIVWE D.

TO
HEMATOLOGY DIVISION, MEDICAL LABORATORY SERVICE
DEPARTMENT, FEDERAL MEDICAL CENTRE ASABA

SUPERVISED BY
MRS EMMANUEL CHINWENDU

23TH MARCH 20231 1

 OUTLINE
 Aim and objective
 Introduction
 Definition
 Genetics
 History
 Epidemiology
 Inheritance pattern
 Pathophysiology
 Clinical features
 Laboratory diagnosis
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 Treatment
 Recommendation
 Conclusion
 Statement

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 AIM
 To understand sickle cell disease and how medical laboratory
diagnosis can detect and help manage its complications.

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 OBJECTIVE
 To understand the processes involved in laboratory diagnosis of sickle
cell disease.

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 INTRODUCTION
 Sickle cell disease (SCD) is a group of blood disorders typically
inherited from a person’s parents.

 It results in an abnormality in the oxygen carrying protein

haemoglobin found in the red blood cells. This leads to a rigid
sickle like shape under certain circumstances.

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 SICKLE CELL ANEMIA
 Sickle cell anaemia occurs when a person inherits two abnormal
copies of the Beta-globin gene leading to a condition in which there
are not enough healthy red cells to carry adequate oxygen
throughout the body

 It is the homozygous state, (HbSS) called sickle cell anemia being

the commonest, other sickle conditions are HbSC, HbSβthal.

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 SICKLE CELL ANEMIA
 In Sickle Cell Anaemia. the red blood cells becomes rigid and sticky
and are shape like sickles or crescent moon.

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 GENETICS
 The change in cell structure arises from a change in the structure of
hemoglobin.
 It is inherited as an autosomal recessive pattern.
 Hb S is caused by a single base change from adenine to thymine in
the coding for the 6th amino acid in the β-chain gene present in the
short arm of chromosome 11.
 Position 6 of normal β-globin chain: Glutamic acid G-A-G.
 Position 6 of Sickle β-chain: Valine G-T-G.

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 EPIDEMIOLOGY
 Sickle Cell Disease is prevalent in areas where malaria is endemic
and in under developed societies.
 People of African descent living outside the continent
 Approximately 8% are carriers and 1 in 375 are born with the
disease.[104]
 HbS gene is found in about 1 in 4 West Africans.
 Nigeria has the highest burden of Sickle Cell Disease with – 30%
of live births having the HbS gene and about – 2% of the
population have HbSS.
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 PREVALENCE
 In 2015, about 4.4 million people have sickle cell disease living in the world
with about 114,800 deaths.[8].
 Mostly descendants of:
 Tropical parts of Africa – East of the Niger River around North Ghana,
Benin, Central African Republic, Nigeria etc.
 Asia – Burma, Thailand,
 Malaysia,
 Indonesia,
 India
 America – LA,
 African American areas Wellems, 2009
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 INHERITANCE PATTERN

 If both parents has sickle cell traits [HbAS] there is a one in four [25%]
chance of having a HbSS offspring

 One in four chance of HbAA

 One in two [50%] chance Of HbAS traits

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 PATHOPHYSIOLOGY
 When HbS is deoxygenated, it polymerises, forming long fibres
(tactoids).
 Becoming rigid, thus deforms the red cell into an elongated and curved
shape called sickles
 Changes can are reversible if the red cell is oxygenated.
 If the deoxygenation persists, the damage becomes permanent and the red
cell is trapped by the reticuloendothelial cells and phagocytosed.
 Red cell lifespan is reduced to about 10-20 days.

Capriotti, 2016

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 PATHOPHYSIOLOGY CONT’D
 Distortion of the red cells causes membrane damage.

 increased K+ efflux accompanied water causing dehydration.

 Sickled cells adher to endothelial cells in the microcirculation.

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 Release of endothelial factors and aggregation of white blood cells and
platelets leading to vasoconstriction and gradual vaso-occlusion.

 In a vicious cycle, hypoxia occurs and hence the sickling.

 Vaso-occlusion leads to ischaemia and subsequently infarction of affected

organs

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HbS Polymer
Vaso-occlusion

Arginine NO

Hemolysis

(Modified from Steinberg, M., Cecil Medicine 2007)

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 CLINICAL FEATURES
 onset of clinical features starts at about 6 months when HbF levels approach
the adult levels.
 Features of chronic haemolysis:
Jaundice,
pallor.
 Steady state chronic haemolysis is usually punctuated by episodes of crises.
There are four types:
1. Vaso-occlusive crisis.
2. Hyper-haemolytic crisis.
3. Visceral sequestration crisis.
4. Aplastic crisis.
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 CLINICAL FEATURES CONT’D
 Features of anaemia that are usually milder in relation to the degree
of anaemia because HbSS has a low affinity for oxygen:
 Lethargy,
 pallor,
 tachycardia,
 tachypnoea,
 dizziness,
 dyspnoea on exertion (DOE),
 dyspnoea at rest (DAR).
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 LABORATORY DIAGNOSIS
 Complete blood count (CBC):
 PCV is usually 15-25%

 T. WBC is mildly elevated (12-16 x 1012/L).

 Platelet count is usually normal or mildly elevated.

 PBF: shows some degree of Anisopoikilocytosis with some fragmented

cells, target cells, sickle cells (more during crisis), polychromasia, features
of hyposlenism eg Howell-Jolly bodies.
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 Screening tests:
 A. Sickling test- the cells in deoxygenated blood sickles when incubated with
reducing agents like Sodium metabisulphite.

 B. Solubility test- HbS is largely insoluble, so it precipitates when incubated

with Sodium dithionite.

 Haemoglobin electrophoresis: separates Hb types based on molecular weight

and electrical charge

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 High performance liquid chromatography (HPLC): based on colour
separation.

 DNA diagnosis: This may be applied to pre-natal diagnosis.

 The samples may be obtained by chorionic villus sampling (CVS), amniotic fluid
sampling.

 The DNA is then analysed using the following techniques

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 DNA DIAGNOSIS
 Polymerase Chain Reaction (PCR):
 Pre-implantation Genetic Diagnosis (PGD) Is Now Possible:
Following IVF, 1 Or 2 Cells Are Removed From The Blastomere
On Day 3. PCR Is Then To Detect The Mutation And Healthy Cells
Are Implanted.

 Foetal Blood Sampling (FBS): Is Done About The Middle Of The

Second Trimester To Obtain Foetal DNA For Study.

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 OTHER ANCILLARY INVESTIGATIONS
 Renal Function Tests:
Urinalysis
Serum Elecrolytes, Urea And Creatinine.

 Chest X-ray (CXR), ECG, Echocardiography, Ventilation-perfusion

Ratio To assess Acute Chest Syndrome (ACS) and Pulmonary
Hypertension.

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 TREATMENT
 Counselling Of Patients And/Or Parents Once Diagnosis Is Made:
Etiology, Course, Investigations, Treatment Options, Financial Burden,
Control Measures.
 Advice On Hygiene And Nutrition.
 Folic Acid Tablets Daily Or Weekly.
 Prophylactic Oral Penicillin Till Puberty.
 Vaccination
Against Encapsulated Organisms Like Haemophilus,
Pneumococcus, Also Meningococcus.
 Crises: Seek Out And Eliminate Precipitating Factors. •
 Nitric oxide.
 Bone marrow transplant.
 Experimental treatment. 29
 TREATMENT CONT’D
 In Pregnancy: Close Attention Is Required Especially Those With A Poor
Obstetric History Who May Need Continuous Transfusion Throughout
Pregnancy.
 Hbs Levels Should Be Reduced To < 30% During Labour.
 Surgery: Close Attention During Anaesthesia And Recovery Ensuring High
Oxygen Saturation And No Acidosis.
 Periodic Transfusions Are Also Advocated In Patients With Repeated
Crises.
 Malaria Prophylaxis In The Tropics.

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 CONCLUSION
 In Conclusion SCA is a inherited red blood cell disorder in which the
cell are distorted and are unable to transport the oxygen carry pigment.

 The faulty hemoglobin is called the hemoglobin S [HbSS].

 SCA is a serious disease that require frequent hospital visit.

 Standardlaboratory investigation is Peripheral blood film and

hemoglobin electrophoresis.
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 RECOMMENDATION
 Genetic Counselling For Intending Couples

 Sickle
Cell Disease Awareness And Campaign ACROSS
COMMUNITIES

 Registration
Of Patients Into Sickle Cell Clinics For Proper
Management And Follow up

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 REFERENCES
 What Is Sickle Cell Disease?". National Heart, Lung, And Blood Institute. June
12, 2015. Archived From The Original On 6 March 2016. Retrieved 8
March 2016.
 Capriotti, Theresa (2016). Pathophysiology : Introductory Concepts And
Clinical Perspectives. Frizzell, Joan Parker. Philadelphia. Isbn
9780803615717. Oclc 900626405.

 Clarke Gm, Higgins Tn (August 2000). "Laboratory Investigation Of

Hemoglobinopathies And Thalassemias: Review And Update". Clinical
Chemistry. 46 (8 Pt 2): 1284–90. Doi:10.1093/Clinchem/46.8.1284.Pmid
10926923. Archived From The Original On 2008-03-20.
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 Gbd 2015 Mortality Causes Of Death Collaborators (October 2016). "Global,
Regional, And National Life Expectancy, All-cause Mortality, And Cause-
specific Mortality For 249 Causes Of Death, 1980-2015: A Systematic
Analysis For The Global Burden Of Disease Study 2015". Lancet. 388
(10053): 1459–1544. Doi:10.1016/S0140-6736(16)31012- 1. Pmc 5388903.
Pmid 27733281.

 Lee, C., Davies, S.,& Dezatoux, C. (2000). Neonatal Screening For Sickle Cell
Disease. The Cochrane Collaboration. John Wiley & Sons, Ltd.

 Wellems TE, Hayton K, Fairhurst RM (September 2009). "The Impact Of

Malaria Parasitism: From Corpuscles To Communities". The Journal Of
Clinical Investigation. 119 (9): 2496–505. Doi:10.1172/Jci38307. Pmc
2735907. Pmid 19729847.
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THANK YOU FOR LISTENING!!!

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