Investigations of blood diseaes
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INVESTIGATIONS OF BLOOD DISEAsES
Presented By: Aayushma Phuyal Roll no: 30 BDS 4th year
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CONTENTSIntroductionCollection of blood sampleHemocytometryInvestigation of blood diseases Red Blood Cell disorder: RBC count RBC indices Mean corpuscular volume (MCV) Mean corpuscular hemoglobin (MCH) Mean corpuscular hemoglobin concentration (MCHC) Mean corpuscular diameter (MCD) Hemoglobin Hematocrit Erythrocyte sedimentation rate (ESR)
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White blood cell disorder Total WBC count Differential count
Test for assessing hemostatis Platelet count Bleeding time Prothrombin time (INR) Von Willebrand’s antigen
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Introduction Hematology is the study of blood. Blood
along with the cardiovascular system performs the
functions of transport, regulation of body
temperature and protection.
It is very useful in the diagnosis and
prognosis of disease
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CLASSIFICATION Disorders involving RBC’s
AnemiaClassification Nutritional deficiency anemia
• Iron deficiency • Pernicious anemia• Folate deficiency anemia
Hemolytic anemia • Thalassemia • Sickle cell anemia• Erythroblastosis fetalis
Aplastic anemia Polycythemia
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Disorders involving WBC’s
Agranulocytosis
Cyclic neutropenia
Lazy leukocyte syndrome
Chédiak-Higashi syndrome
Infectious mononucleosis
Leukemia
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Disorders involving plateletsPurpura
Diseases involving specific blood factors
Hemophilia
Von Willebrand’s disease
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Collection of blood sample
Asepsis
Condition of being free from septic or infectious
material – bacteria, viruses etc
Sterilization of equipment
Syringes, needles, lancets, cotton and gauze swabs
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Sources and amount of blood sample
Blood sample
Capillary blood
Venous blood
Arterial blood
Cardiac catheterization
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HEMOCYTOMETRY
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Procedure of counting the number of cells in a
sample of blood, the red cells, the white cells,
and the platelets being counted separately.
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Principle The sample of blood is diluted in a special pipette and
placed in a capillary space of known volume between a
specially ruled glass slide and a cover slip
The cells spread out in a single layer makes counting easy
Knowing the dilution employed, the number of cells in
undiluted blood can be easily calculated
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Hemocytometer
Consists of the following
Diluting pipettes
Also called “cell pipettes” or “blood pipettes”
RBC and WBC pipettes
Counting chamber
Coverslips
RBC and WBC diluting fluids
Watch glasses, spirit swabs, blood lancet etc
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Hemocytometer
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The Total WBC Count
Principle
A sample of blood is diluted with a diluting fluid
which destroys the red cells and stains the nuclei
of the WBC.
The cells are then counted in a counting chamber
and their number in undiluted blood reported as
leucocytes/mm3
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Normal leucocyte count: 4000 – 11,000 cells/mm3
Leucocytosis
Increase in WBC count beyond 11,000 cells/mm3
Physiological leucocytosis
Infants
Pregnancy
Physical exercise
Mental stress
Parturition
Extremes of temperature
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Pathological leucocytosis
Acute infection with pyogenic bacteria
Myocardial infarction
Acute hemorrhage
Burns
Malignancies
Surgical operations
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Leucopenia
Decrease in the number of white cells below the 4000/mm3
level
Pathological leucopenia
Infection with non pyogenic organisms
Viral infections
Drugs
Repeated exposure to X-rays and radium
Malnutrition
Hypoplasia and aplasia
Hemopoetic disorders
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Differential leukocyte count
• Leishman stain• Wright stain• Giemsa• May Grunwald Giemsa stain
Principle
A blood film is stained with Leishman’s stain and scanned
under oil immersion.
The different types of leukocytes are identified and their
percentage distribution is calculated.
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Ideal film thickness
Thin blood film Thick blood film
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Agranulocytes
Granulocytes
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WBC count
Increased Decreased
Leukemia
Infectious mononucleosis
Leukemoid reaction
Agranulocytosis
Cyclic neutropenia
( < 2000 cells/ mm3 )
Rhythmic3 week cycles
Normal blood count (4-5 days)↓
Decreased neutrophil count (increase in mono & lymphocytes; peak – completely disappear)
↓Cells reappear in 4-5 days
↓Normal blood count
absolute neutrophil count
remaining above 30,000/mm3
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Acute leukemia
WBC > 1,00,000/cc
Increased BT and CT
Chronic leukemia
WBC > 5,00,000/cc
Anemia
Thrombocytopenia
Myeloblast/ undifferentiated myelocyte
Stem cell leukemia - undifferentiated
Pictures Of Blood
Normal human blood
White Cell Red Cell
Platelet
Blood with leukemia
BlastsRed Cell
Platelet
White Cell
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WBC count
Qualitative defects Quantitative defects
Lazy leucocyte syndrome
Chédiak Higashi Syndrome
Agranulocytosis
Cyclic neutropenia
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Laboratory findings
Lymphocytosis (Increased WBC’s)
Atypical lymphocytes (horseshoe shaped or
indented nucleus, foamy cytoplasm)
Antibodies to EBV
Paul-Bunnell test
Monospot test
Infectious mononucleosis
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Paul Bunnel test (serological test)
Heterophile test
Ig M antibodies elicited by EBV infection
Acute phase – 80 -90% exhibit these Abs
Property - Agglutinate sheep erythrocytes
Titre of agglutinins & hemolysins against sheep RBC increased
4th week – titer decreases 3 month – disappears
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Paul Bunnel test (serological test)
Serum + equal volume of 1% sheep erythrocyte suspensionIncubated at 370 - 4 hrs
Examined for agglutination
Infectious mononucleosis – > 100 increase in the titre Normal serum – 1:8
1:4096
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Monospot test
Commercially available
Rapid qualitative slide agglutination test
Screening tool
Agglutination of horse RBCs on exposure to EBV heterophile
AB
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Tests for Hemostasis
It is a homeostatic mechanism to prevent loss of
blood.
Hemostasis involves the following steps
1. Vaosconstriction
2. Platelet plug formation
3. Formation of blood clot
4. Fibrinolysis
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Bleeding time
It is the interval between skin puncture and
spontaneous unassisted stoppage of bleeding
It is an in vitro test of platelet function
• DUKE METHOD (1-5 min)
• IVY METHOD (9 min)
• SIMPLATE METHOD (>7 min)
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Normal bleeding time : 1- 5 mins
Advantages Simple and quite reliable
Duke bleeding time
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Ivy bleeding time
Method is more reliable than Duke’s methodNormal bleeding time with this method: 9 mins
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Thrombocytopenia
1. Decreased production
Bone marrow injury or failure or depression
Bone marrow invasion
2. Increased destruction
Drugs
Immune thrombocytopenic purpura
Sequestration in spleen
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Clotting time
Capillary blood clotting time:
Also called Wright’s capillary glass tube method
Results:Normal clotting time: 3 – 6 mins
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Drop method
Less accurate than the Wright’s capillary glass tube method.
• Normal clotting time : 2 – 4 mins
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Venous blood clotting time Also called Lee and White test tube methoda. Single test tube method b. Most widely used method c. Normal clotting time : 5 – 10 mins
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Increase in CT•Hereditary coagulation disorders•Haemophilias•von Willebrand disease•Afibrinogenemia and dysfibrinogenemia•Deficiency of factor XIII •Acquired coagulation disorders•Vit K deficiency•Liver diseases•Intravascular clotting•Anticoagulant therapy•Newborns
Decrease in CT•Physiological conditions•Malnutrition•Parturition
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Disease BT/CT Reason Thrombocytopenic Purpura BT-Prolonged Abnormal reduction in circulating
platelets
Familial Thrombasthenia BT Prolonged Qualitative defect in plateletPlatelet count is normal
Thrombocytopathic Purpura BT Prolonged Qualitative defect in plateletPlatelet count is normal
Thrombocythemia BT Prolonged Great increase in the platelet count – interferes with formation of thromboplastin
Abnormal aggregating agents
Von Willebrand’s Disease CT – ProlongedBT – Prolonged
Abnormality of von Willebrand factor
Parahemophilia CT – Prolonged BT – Normal
Reduction in plasma Proaccelerin (factor V)
Afibrinogenemia and Hypofibrinogenemia
CT – Prolonged BT – Normal
Little or no fibrinogen present in tissues or plasma
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INR = PT test PT standard
•ISI: international sensitivity index (analytical system)•Normal value = 1 Patients on anticoagulants: 2-3
Measures extrinsic pathway activity•Prolonged Prothrombin time may be due to:• Oral anticoagulants.• Deficiency of factor I, II, V, VII or X•High levels of heparin
Activated partial thromboplastin time (aPTT)•Measures the intrinsic pathway of coagulation•Prolonged APTT •Deficiency of factors I, II, V, VIII, IX, X, XI, or XII•Heparin therapy, inhibitors to specific coagulation factors•High levels of FDPs (fibrinogen degradation products)
ISI
International Normalised Ratio (INR)
Prothrombin Time
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SUMMARYBleeding Time: Duke’s method=1-5 min
Ivy method=9 min
Simplate method =>7 min
Clotting Time: Wright’s method=3-6 min
Drop method= 2-4 min
Duke’s method= 1-5 min
Lee & White method=5-10 min
The average value of BT &CT is taken into consideration while doing any
investigations of the blood diseases, any fluctuation in these values
may alter the treatment plan.
Hence these values are important to rule out various
disorders and it’s needful treatment.
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• Routine blood investigations are an inherent part of pathology.
• These investigations many-a-times may be diagnostic and/or
adjunct to diagnosis for various pathologies.
• Both the peripheral blood smear and hemoglobin concentration
gives us important diagnostic clues, so understanding and
reading these basic hematological techniques will helps us to
provide a better diagnosis.
CONCLUSION
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REFERENCES
ESSENTIAL OF PHYSIOLOGY BY: ABS MAHAPATRA(3rd edition)BURKET’S ORAL MEDICINE BY: GREENBERG GLICK SHIP (11TH EDITION)
SHAFER’S ORAL PATHOLOGY: (7TH EDITION)
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