This document summarizes hemolytic anemia, including its causes, characteristics, laboratory tests used for evaluation, and specific types like hereditary spherocytosis, hereditary elliptocytosis, G6PD deficiency, sickle cell disease, and thalassemia. Key tests discussed are serum bilirubin, urine urobilinogen, fecal stercobilinogen, serum haptoglobin, LDH, reticulocyte count, and the Coombs test for immunohemolytic anemia. Causes of hemolytic anemia include blood loss, increased red blood cell destruction (hemolysis), and increased red blood cell production.

1. INVESTIGATION OF HAEMOLYTIC
ANAEMIA
BY,
MOHANKUMAR.M
43RD VZ

2. ANEMIA
 Quantitative and qualitative reduction of RBCs leading
to low oxygen carrying capacity of the blood
irrespective of age & sex.
 Normal Hb:
 Males: 14-18 gm/dl
 Females:12-15.5 gm/dl

3. CLASSIFICATION OF ANEMIA
 Blood loss
 Due to increased red cell destruction-HEMOLYSIS
 Due to increased red cell production

4. HEMOLYTIC ANEMIA
 Defined as anemia's resulting from an increase in the
rate of red cell destruction.
 INTRAVASCULAR HEMOLYSIS-Red cells undergo
lysis in the circulation and release their contents into
plasma.
 EXTRAVASCULAR HEMOLYSIS-Red cells are taken
up by the cells of the RE system where they are
destroyed and digested.

5. CHARACTERISTICS
 Anaemia
 Jaundice.
 Splenomegaly.
 Increased reticulocytes i.e.. Brisk reticulocytosis.

6. LIST OF HEMOLYTIC ANEMIAS
 Hereditary spherocytosis
 Hereditary elliptocytosis
 Sickle cell disease
 Thalassemia
 Hemolytic anemia due to G6PD deficiency
 Immunohemolytic anemia
 Paroxysmal nocturnal hemoglobinuria

7. RBC INDICES
 Mean Corpuscular Volume
MCV=PCV per 100 ml blood/RBC count in
million per µL.
Normal=78-94 fL.

8.  Mean Corpuscular Hemoglobin
MCH=Hemoglobin in gm per dL/RBC count in
million per µL.
Normal=28-32 pg.

9.  Mean Corpuscular Hemoglobin Concentration
MCHC=Hb in gm per dL/PCV per 100 ml blood
Normal=30-37%

10. LABORATORY EVALUATION OF
HAEMOLYSIS
A. Test of increased red cell breakdown
1.Serum bilirubin-unconjugated bilirubin is
increased (van den Bergh reaction-indirect
positive)
2.Urine urobilinogen is raised.
3.Faecal stercobilinogen is raised.
4.Serum haptoglobin(α-globulin binding protein)
is reduced or absent.
5.Plasma lactate dehydrogenase is raised.

11. B.Test for increased red cell
production
1.Reticulocyte count reveals reticulocytosis which is
generally early and is hence most useful initial test of
marrow erythroid hyperplasia.
2.Routine blood film(PS) shows macrocytosis,
polychromasia and presence of normoblasts.
3.Bone marrow shows erythroid hyperplasia with usually
raised iron stores.
4.X-ray of bones shows evidence of expansion of marrow
space, especially in tubular bones and skull.

12. Hemolytic anemia

14. Hereditary spherocytosis

15. Hereditary elliptocytosis

16. G6PD deficiency

17. Sickle cell disease

18. Sickle cell disease

19. Thalassemia

20. Thalassemia

21. Microangiopathic hemolytic
anemia

22. C.Test of Damage to Red Cell
1.Routine blood film.
2.Osmotic fragility test.
3.Coomb’s antiglobulin test.
4.Eletrophoresis for abnormal hemoglobin
5.Estimation of HbA2.
6.Estimation of HbF.
7.Tests for sickling.
8.Screening test for G6PD deficiency.

23. D.Tests for Shortened Red Cell Life
Span
A shortened red cell survival is best tested by 51Cr
labeling method. Normal RBC life span of 120 is
shortened to 20-40 days in moderate haemolysis and
to 5-20 days in severe haemolysis.

24. E.OTHER TESTS-
FLOWCYTOMETRTY

26. IMMUNOHEMOLYTIC ANEMIA-
COOMB’S TEST
Immunohemolytic anemia's are a group of
anemia's occurring due to antibody production by the
body against its own red cells. It is induced by one of
the following three types of antibodies:
1.Autoimmune hemolytic anemia
a. Warm antibody AIHA
b. Cold antibody AIHA
2.Drug induced immunohemolytic anemia
3.Isoimmune hemolytic anemia

27. IMMUNOHEMOLYTIC ANEMIA-
COOMB’S TEST (Contd.)
 An important diagnostic tool in all cases of
immunohemolytic anemia's is Coomb’s antiglobulin
test.
 It is the test for detection of Rh-antibodies in
saline(direct Coomb’s) or after addition of
albumin(indirect Coomb’s).

31. Electrophoresis picture

32. SICKLING TEST
 It is done under condition of reduced oxygen tension
by an oxygen consuming reagent, sodium
metabisulfate.
 It is a confirmatory test for sickle cell disease.