1. Investigation in hematology
Chittima Sirijerachai

2. Investigation in hematology
• Anemia
• Bleeding
• Malignancy

3. Investigation in patient with anemia
Specific
Initial
• Inclusion body
• CBC, blood smear
• Osmotic fragility
• Red cell indicies • Saline agglutination
• Coombs’ test
• Reticulocyte count
• Hb typing
• Ham’s test
• BM aspiration

4. CBC
-anemia
Hb, HCT
-polycythemia
-leukocytosis
WBC
-leukopenia
-abnormal cells
-thrombocytosis
Platelet
-thrombocytopenia

5. Criteria for Diagnosis Anemia
Hb.( g/dl) Hct (%)
Children 11.0 33
Male 13.0 40
Female ( 15-50 ) 12.0 36
Female ( > 50 ) 13.0 40
Pregnant women 11.0 33

6. Criteria for diagnosis polycythemia
Hb Hct
• Male 18 60
• Female 17.5 55

7. CBC
-anemia
Hb, HCT
-polycythemia
-leukocytosis
WBC
-leukopenia
-abnormal cells
-thrombocytosis
Platelet
-thrombocytopenia

8. Normal value
WBC 5,000 – 10,000 /cu.mm.
PMN 40-75 %
L 20-50 %
Eo 1-6 %
B 1 %

9. Leukocyte disorders
leukocytosis leukopenia
neutrophilia neutropenia
lymphocytosis lymphopenia
eosinophilia
basophilia
monocytosis
leukemoid reaction
leukemia
leukoerythroblastic (myelophthisis)

15. CBC
-anemia
Hb, HCT
-polycythemia
-leukocytosis
WBC
-leukopenia
-abnormal cells
-thrombocytosis
Platelet
-thrombocytopenia

17. CBC and RBC indices
80-96 fl.
27-33 pg.
33-36 g/dl
12.0-13.6%

18. Reticulocyte count
Normal = 0.2-2 %
Corrected reticulocyte = Pt HCT X Reti.
Normal HCT

19. Inclusion body
Heinz body

20. Osmotic fragility (OF)
• Red cell + 0.33 % NSS
• One tube OF – screening for thalassemia
• Hereditary spherocytosis

21. Bone marrow aspiration
Indication :-
• Cytopenia
• Myelophthisis anemia
• Leukemia
• Lymphoma staging
• Myeloproliferative disorders
• Fever of unknown origin

22. Thalassemia
- α thalassemia
- β thalassemia
- hemoglobinopathy

23. α thalassemia
α1 trait = _ _ /αα
α2 trait = _ α/αα
α1 α2 = __/_α
α1 α1 = __/__
α 2 α2 = _ α/ _ α

24. β thalassemia
β 0
β +

25. Normal hemoglobin
α 2β 2 = Hb A
β
α α 2γ 2 = Hb F
γ α2 δ2 = Hb A2
δ

26. α thalassemia
Hb A
β Hb A2
Hb F
α γ
δ
β4 = Hb H
γ4 = Hb Bart’s

27. β thalassemia
Hb A
α β
Hb A2
γ Hb F
δ

28. Heterozygote hemoglobin E
β α 2β 2 = Hb A
Α
α βE
α 2β Ε
= Hb E
γ 2
δ 25-35 %

29. β thalassemia/Hb E
Hb A / absent
α β E Hb A2/ Hb E
γ
Hb F
δ

30. Father Mother sibling 1 sibling 2
Hct 45% 33 % 42% 36 %
MCV 70 65 82 70%
Hb typing A2A A2ABart’s H A2A A2A
Hb A % 96 91 97 98
Hb A2 2.2 1.4 1.1 1.5
Hb F 1.3
Hb E
Hb Bart’s 3.3
Hb H 7.5
Hb CS
α 1 trait α 1α 2 α 2 trait α 1 trait
genotype

31. Father mother sibling 1 sibling 2
Hct 40 % 40% 25 45%
MCV 72 84 65 85
Hb typing A2A EA EF A2A
Hb A % 94 98
Hb A2 5.5 1.5
Hb F 32
Hb E 30 65
Hb Bart’s
Hb H
Hb CS
β trait E trait β /E normal
genotype

32. Investigation in patient with
bleeding disorders

33. Investigation
CBC, platelet count
Bleeding time
Platelet function test
Venous clotting time
Prothrombin time (PT)
Partial thromboplastin time (PTT)

34. CBC and platelet
Platelet: < 20,000 Spontaneous bleeding
20,000-50,000 Bleed after minor trauma
50,000-70,000 Bleed after major trauma

36. CBC and platelet

37. Giant platelet

39. Bleeding time
•Platelet function
•Normal: 2-7 minute

41. PTT

42. PT

43. TT

44. Venous clotting time
Normal: 5-20 minute
Prolong if clotting factor is <5 %
Small clot - hypofribinogenemia
Clot retraction time (plt function)
Clot lysis time

45. Prothrombin time (PT)
Normal: 11.6-14.4 sec
PT ratio = patient PT Normal < 1.3
Control PT
ISI
INR = patient PT
Control PT
ISI = international sensitivity index

46. Partial thromboplastin time (PTT)
Normal = 34-48 sec
Isolated PTT prolong:
•F VIII, IX, XI, XII deficiency
•F VIII, IX, XI inhibitor
•Antiphospholipid syndrome

47. Mixing test
Patient plasma 0 hour
PTT
Normal plasma 2 hour
Correctable PTT Factor deficiency