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1. The Role of Lab Exam. Screening Diagnosis : Routine Lab tests Confirmatory Lab tests Prognosis Monitoring Disease activity Therapy responses. 2. Laboratory examination for Infection. 3. Routine examination. HEMATOLOGY :. Blood cell count complete blood cont (CBC)
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The Role of Lab Exam • Screening • Diagnosis : • Routine Lab tests • Confirmatory Lab tests • Prognosis • Monitoring • Disease activity • Therapy responses 2
Routine examination HEMATOLOGY : Blood cell count complete blood cont (CBC) • Hemoglobin concentration (Hb) • White Blood Cell Count (WBC) • Platelet count • Differential cell count • Red blood cell count & Hematocrit Erythrocyte Sedimentation Rate (ESR) 4
Routine examination - hematology • Hemoglobin concentration • Normal range : • At birth : 15 – 20 g/dl • At 2 months : 9 – 14 g/dl • 10 years of age : 12 – 15 g/dl • Female adult : 12 - 16 g/dl • Male adult : 13 – 18 g/dl • < Normal range : Anemia • Anemia occur in several infection diseases as follows: - bacterial infection • - virus infection • - parasite infection Blood cell count 5
Anemia in bacterial infection Extracellular microorganism Clostridial Septicemia Bartonellosis Invade to RBCs Adhere to the exterior surface of the RBC Destruction of RBCs Lysis ANEMIA 6
Hemolytic anemia in parasites infection Infected cell Immune complexes ruptures Lysis ANEMIA 7
Anemia of Chronic Disease ACD is associated with an underlying disease (usually inflammation, infection, or malignancy), but is without apparent cause (not due to a lack of the nutrients iron, vitamin B 12, or folic acid) Anemia of chronic disease (ACD) is difficult to define as its etiology and pathogenesis is not clear. ACD is the most common anemia in hospitalized patients. 8
Anemia of Chronic Disease Pathophysiology: • Erythropoesis suppression • Chronic inflammatory process secretion of TNF & IL-1 • Lack of iron for Hb synthesis • Lactoferrin release from granules of neutrophils • Lactoferrin competes with transferrin for iron • Decreased RBC survival 9
Routine examination - hematology LEUKOCYTE COUNT (WBC) • Measure number of total leucocytes • Method: manually & automatically • Principle : dilution of blood with acid solution in order to lyses erythrocytes • Reference range : adult = 4000 -11.000 cells/μL child = 4500-17.000 cells/μL newborn= 6000-30.000 cells/μL 10
Kinetics of Leucocyte Storage pool Circulating pool Marginal pool Input from marrow Output to tissue 11
Pathology WBC Virus infection Typhoid fever Rheumatoid arthritis Cirrhosis of the liver SLE Radiation, drugs Bacterial infection Leukemia Uremia Physiologic: Pregnancy Strenuous exercise Emotional stress, anxiety Leukocytosis WBC > 11.0 (x 109/L) Leukopenia WBC < 4.0 (x 109/L) 12
Routine examination - hematology White Blood Cell Differential • To determine the relative number of each type of WBC present in the blood. • Blood smear : - relative number - leukocyte immaturity - morphologic abnormality • Abnormality: Quantitative Qualitative 13
Classification of Leucocytes Granulocyte Neutrophil, Eosinofphl, Basophil Polimorfonuclear Neutrophil, Eosinofphl, Basophil Phagocyte Neutrophil Monocyte Non-granulocyte Monocyte Lymphocyte Mononuclear Monocyte Lymphocyte Immunocyte Lymphocyte 14
All white blood cells originate from the bone marrow Growth and differentiation factors (cytokines) produced by and present on bone marrow stromal cells determine the type of white blood cell that will emerge, as well as their relative numbers. 15
Blood cells migrate through blood and lymph nodes or home to tissues 17
Cells in blood circulation Very few in blood 19
Differential cell count • Polymorphonuclear neutrophils : 50 – 70 % • Bands : 0 – 5 % • Lymphocytes : 18 – 42 % • Monocytes : 1 – 10 % • Eosinophils : 1 – 4 % • Basophils : 0 – 2 % • Course of d’s : shift to the left (acute), shift to the right (chronic) • Cause : bacterial, viral and parasites infection • neutrophilia (bacterial infection), lymphocytosis • (viral infection, tuberculosis) Refference range: 21
NEUTROPHILIA 3 major cause : infection, inflammation, malignancy Severity of neutrophilia in infection depend on: - virulency of organism, - age : child > - patient immunity: immunocompromised host Quantitative abnormality 22
Causes of neutrophilia Quantitative abnormality 1. Bacterial Infection 2. Toxic agent 3. Metabolic: uremia, eclampsy, metabolic acidosis 4. Drugs & chemicals: mercury, digitalis, steroid 5. Physic & emotional stimuli 6. Tissue damage & necrosis: myocardial infarct, wound, neoplastic diseases 7. Hemorrhage: especially intra serous cavity (peritoneal, pleural, joint space, subdural) 8. Hematological diseases: leukemia. 23
Qualitative Abnormality Shift to the left or right: promielosit mielosit metamielosit mieloblas batang segmen • Shift to the left : • increase immatur cells • most frequent: stab, • metamielosit, mielosit, promielosit • acute infection (bacterial) • Shift to the right: • increase of segment • hypersegmentation • chronic infection 24
Leukemoid reaction mielocytic/netrophyilic Quanti+Qualitative abnormality 25 Bain, 2002. Blood Cells, A Practical Guide,3rd ed, Blackwell Publ, UK
Qualitative abnormality White blood cell (blood smear) vacuolisation vacuolisation Toxic granulation Leucocytosis : netrophilia absolute with toxic granulation & vacuolisation Bacterial infection 26
Vacuolisation & toxic granulation Toxic Granulation vakuolisation Bacterial infection 27
Qualitative abnormality • Toxic Granulation • Stimulated by organism or antigen • Color of granule: dark blue-blackish • Profound toxic granulation worse prognosis • Vacuolisation of cytoplasm phagocytosis process 28
Neutropenia Netropenia lekopenia Agranulositosis: severe netropenia Causes of netropenia: Viral infection Certain Bacteria: Tifoid/ paratifoid Severe infection Immune reaction: autoimmune/ drug induced 29
EOSINOPHILIA : 1. Parasite investation - correlate with killed parasites - eosinophyl attracted to parasite will be killed by degranulation process 2. Allergy/ hypersensitivity 30
Lymphocytosis Absolute lymphocytosis Viral infection 32
Qualitative abnormality Variant / atypical/ virocyte/ reactive lymphocyte response to infection 33
Lymphocytosis with variant lymph: - Mononukleosis infecsiosa (var lymph 40%), acute hepatitis, citomegalovirus (CMV) - measles, pneumonia viral, rubela relatif - Non viral : Tuberculosis, syphilis, malaria, typhus, diphteria, toxoplasmosis • Lymphocytosis without var lymph: asimptomatic viral inf., diarrhea, resp. inf • Lymphopenia; HIV, SLE, intensive chemotherapy 34
Virus Infection MONONUKLEOSIS INFEKSIOSA (MI) cause: virus Epstein-Barr (EBV) Lekositosis with limphocytosis, dan atypical lymphocyte “Kissing-cell” 35
Dengue virus infection Reactive Lymphocyte Blue cytoplasm- Lymphocyte 36
Monocyte 37
MONOCYTOSIS • Some bacterial inf.,: - Active Tuberculosis : - Sub acute bacterial endocarditis - Syphilis • Myeloproliferatif • Recovery 38
Erythrocyte Sedimentation rate(ESR) Routine examination - hematology ESR is the rate in millimeters at which the RBCs fall in 1 hour Monitoring the course of an existing inflammatory disease Normal range: 0-20 mm/hrs F 0-15 mm/hrs M Elevated : bacterial infection 39
Normal sedimentation • Polisitemia : AE • Dekompensasi jantung • Sickle sel anemia, sferositosis • Neonatus Increase Sedimentation • infection • myocardial infarct • Rheumatic fever • Malignancy with necrosis • Active tuberculosis , tissue destruction • Surgery Trauma, shock • Hiperglobulinemia • Pregnancy 40
C-REACTIVE PROTEIN (CRP) • an acute phase reactant • In general parallel ESR but not influenced by erythrocyte • More sensitive than ESR • Increase & decrease faster : - early indicator of acute infection - monitor course of disease 41
CRP increase in : • Infection: • Lower in viral compared to bacterial infection • Useful to monitor disease activity • Inflammatory disorders: • Earlier,more intense increase than ESR • Dissaperance of CRP precedes the return to normal of ESR • Tissue injury or necrosis • AMI : appears within 24-48 hrs • Malignant disease, Following surgery, burns 42