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1. Full blood count
2. Hb
3. Blood picture
4. Bone marrow investigations
5. Reticulocyte count
6. Serum iron studies
7. Eletrophoresis
8. Coombs test
9. Osmotic fragility test
10.ESR
11.Serum creatinine levels
1.RBC count
2.Hematocrit
3.MCV
4.MCH
5.MCHC
6.RDW
 Iron deficiency anaemia
 Thalasaemia
 Sideroblastic anaemia
SERUM IRON LEVELS Significantly decrease
TOTAL IRON BINDING Increased
CAPACITY
SERUM FERRITIN Decreased
LEVEL
TRANSFERRIN Significantly decreased
SATURATION
 Alpha-Thalassemia
 Beta-Thalassemia Thalassemia major
Thalassaemia trait
Low MCV
Blood film
Serum iron
Normal/High
Hb F/A2
High Low
Bone marrow
iron
Sideroblastic
anaemia
Ferritin
Iron deficiency
anaemia
Thalassaemia
Macrocytic
anaemia
Megaloblastic
anaemia
Non megaloblastic
anaemia
Vit B12
dificiency
anaemia
Folate
dificiency
anaemia
Alcoholic
Chonic liver
disaese
dugs
Hypothyroidism Aplastic
anaemia
Vit B12 Dificiency aneamia
We can do further
investigations to confirm the
cause
Intrinsic factor antibody test
Schilling test
Upper gastro intestinal endoscopy
History taken
Folate Dificiency anaemia
Red cell folate test
Serum folate test
Some of main causes are
Excess alcohol
Liver disease
Aplastic anaemia
Hypothyroidism
Drugs
This causes can confirmed
by History taken of the
patient
High
MCV/MCH
Blood film
Reticulocyte
count
Normal or
low
Non
megaloblastic
Bone
marrow
morpology
Megaloblastic
Folate
and vit
B12
Folate
low
B 12 low
Folate B 12
Normoblastic Dyserypoietic
MyelodysAlcoholicChronic liver
disease
Normal
MCV/MCH
Blood film
Reticulocyte
count
High Normal or low
Bone marrow
morpology
HemolysisAcute blood
loss
Increased RBC production Increased RBC break down
Reticulocytosis
serum bilirubin
LDH
Urobilinogen
+ ve haemosiderin
Plasma haptoglobulin
Confirm hemolytic anemia
 Autoimmune hemolytic disease
 Hereditary spherocytosis
 G6PD deficiency
 Sickle cell anaemia
 Non autoimmune-microangeopathic hemolytic
anaemia
2. osmotic fragility test
Hereditary spherocytosis
patient
Normal blood picture
BloodPicture
Bite or Blister cells
1.Hemoglobin concentration < 10 g/dL, or
Absolute reticulocyte count < 40,000/μL (40
× 109/L).
2. Total leucocyte count <4000/μL(4x109/L)
Absolute Neutrophil count < 1500/μL (1.5 ×
109/L),
3.Platelet count < 150,000/μL (150 × 109/L)
PANCYTOPENIA AT INITIAL
EVALUATION
REFER PATIENT FOR URGENT
EVALUATION
REPEAT CBC AND BLOOD SMEAR
BONE MARROW ASPIRATE
ANDTREPHINE BIOPSY
BONE MARROW CYTOGENETICS
BONE MARROW
IMMUNOPHENOTYPING
LFT,B12 AND FOLATE,COAGULATION
PROFILE,VIRAL
ETIOLOGY,AUTOIMMUNE PROFILE
1. APLASTIC ANEMIA
2. HYPOPLASTIC
MYELODYSPLASTIC SYNDROME
3. LYMPHOMA IN HYPOPLASTIC
BONE MARROW
4. CYTOTOXIC AGENTS AND
RADIOTHERAPY
VERY RARE- ACUTE LEUKEMIA IN
HYPOPLASTIC BONE MARROW
1. ACUTE LEUKEMIA/
LYMPHOMA
2. HAIRY CELL LEUKEMIA
3. MYELOFIBROSIS
4. MYELODYSPLASTIC
LEUKEMIAS
5. PAROXYSMAL NOCTURNAL
HAEMOGLOBINURIA
6. MULTIPLE MYELOMA
1. METASTATIC SOLID TUMOURS
2. HYPERSPLENISM
3. DEFICIENCY OF VITAMIN B12 AND
FOLIC ACID
4. INFECTIONS: -SEPSIS
- TB,KALA AZAR,BRUCELLOSIS
5. OVERWHELMING INFECTIONS
6. ALCOHOL
7. AUTOIMMUNE DISORDERS- SLE,
SJOGREN’S
8. SARCOIDOSIS
Normal distribution and
cellularity are seen
Normal bone marrow
biopsy
Aplastic patient
a – bone marrow aspiration
a ̶ bone marrow biopsy
Hypocellularity can be seen
Aplastic anaemia
TEST RATIONALE
BONE X-RAYS Multiple myeloma, metastasis.
BLOOD CULTURE Infectious agent- Tuberculosis or virus.
VITAMIN B12 AND FOLATE ASSAYS Megaloblastic anemia
ASPARTATE AMINOTRANSFERASE,
ALANINE AMINOTRANSFERASE,
GAMMA GLUTAMYL
TRANSFERASE, BILIRUBIN
Evaluate hepatitis
BLOOD UREA NITROGEN,
CREATININE
Assess for Chronic Renal Failure
SEROLOGY For HIV, EBV, Hepatitis
HAM’S TEST Paroxysmal Nocturnal Haemoglubinuria
CHROMOSOMAL BREAKAGE
STUDIES
Fanconi anemia
 Classification :
 Acute
 Acute lymphoblastic leukemia (T-ALL & B-ALL)
 Acute myeloid leukemia
 Chronic
 Chronic myeloid leukemia
 Chronic lymphocytic leukemia
1. Full blood count
 reduced
haemoglobin
normochromic,
normocytic anaemia,
 WBC
<1.0x109/l to >200x109/l,
neutropenia and f
blast cells
 Thrombocytopenia
 <10x109/l).
Acute lymphoblastic
leukemia
Acute myeloid leukemia
 ALL(Lymphoblast)
 Blast size :small
 Cytoplasm: Scant
 Chromatin: Dense
 Nucleoli :Indistinct
 Auer-rods: Never
present
 AML (Myeloblast)
 Large
 Moderate
 Fine, Lacy
 Prominent
 Present in 50%
2. Bone marrow
aspiration and
trephine biopsy
 confirm acute
leukaemia
(blast > 30%)
 usually
hypercellular
3. Cytochemical
staining
a) Peroxidase :-
 * negative ALL
 * positive AML
Positive for myeloblast
b) Periodic acid
schiff
*Positive ALL
(block)
* Negative AML
Block positive in ALL
c) Acid phosphatase :
focal positive
(T-ALL)
 Monoclonal antibodies(McAb) are recognised
under a cluster of differentiation(CD).
MONOCLONAL ANTIBODIES USED FOR
CHARACTERISATION OF ALL AND AML.
Monoclonal antibodies
AML : CD13, CD33
ALL : B-ALL CD10, CD 19, CD22
T-ALL CD3, CD7
Anoop k r

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