‫شادي الحربي‬ ‫ملخص علم الدم‬

‫بسم هللا الرحمن الرحيم‬

Hematopoiesis
-It is Formation of different blood cells (RBCs-WBCs-PLATELATE).
- Site: Red Bone Marrow in all bone of children (Yolk sac), But in adult restricted to
Central Skeleton (skull, sternum. rib, hips, and vertebra).
* Factors Necessary For Erythropoiesis (Formation of RBCs):-
1- Erythropoietin hormone. 2- Iron
3- Vit B12 4- Folic Acid
5- Ascorbic Acid (Vit C) 6- Vit B6 (Pyridoxine)
7- Amino Acid.
* RBCs: Red color, un nucleated, biconcave shape.
-About 1% of RBCs per day are Breakdown
Function of RBCs:
Transport Oxygen + CO2 + Hormone + Enzyme +Vitamins + Nutrients
*Reticulocytes:-
-It is immature RBCs.N.R 0.5-3% and stained by Brilliant Cresyl Blue.
Reticulocytosis:
1-hemorrhage. 2-megalobastic anemia 3-hemolytic anemia.
Reticulocytopenia:
1-B.M suppression (by drugs). 2- B.M failure (aplastic anemia).
Erythrocytosis: (Increase in red Blood Cells with physiological causes):-
1- High altitudes. 2- Physical training.
3- Drugs. 4- Smokers.
Polycythemia:
-It is increased RBCs count, Hb, Hct, and PCV. With Pathological causes.
-The Increase of RBCs is not by physiological causes.
Types of it:
1-Primary polycythemia. 2-Secondary polycythemia.
3-Relative polycythemia.
Relative polycythemia:
-It is due to decreased plasma volume with normal RBCs count.
– Relative increased of PCV but normal WBCs and Platelets.
e.g: Burns, diuretic ttt
Erythrocytopenia: (Decrease in Red Blood Cells)
1- Hemorrhage 2- Abnormal destruction of RBCs
3- Lack substance needed for RBCs Production.
4- Chemotherapy or radiation 5- Leukemia.

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 ‫شادي الحربي‬ ‫ملخص علم الدم‬

*Parameters of CBC (Complete Blood Count):-

- Hemoglobin: Hb(13-17 g\dl)
- Hematocrite: HCT (40-50% )
- Mean cell volume: MCV (80 – 100 FL)
- Mean cell hemoglobin weight: MCH (27– 33 pg)
- Mean cell hemoglobin concentration: MCHC (32 – 36 %)
* Alteration in size of RBCs:- anisocytosis.
* Alteration in shape of RBCs: Poikilocytosis
*Hemoglobin: Consist Of 4 Protein (94%) + 4 Heme groups (6%)

*Leucocytosis: increased in WBCs count.

*Neutrophilia: increased in neutrophil count
Causes:-
-physiological:
1-Pregnancy 2- after exercise 3- lactation. 4- Neonates (new borne)
-pathological:
1- acute pyogenic infection (Bacterial) 2- hemorrhage or hemolysis
4- CML 5- polycythemia rubra vira
3- Trauma or surgery 6- Dohle bodies seen
*Eosinphilia: increased in eosinophil count.
Causes:
1- Allergy 2- parasitic infestation
3- Steven's–johnson syndrome. 4- Dermatitis
5- Loffler's syndrome 6- Hodgkin's disease
*Lymphocytosis: increased in lymphocytic count
Causes:-
-Physiologic: 1-infants 2-children
-Pathological:
1- Viral infection 2- Some bacterial infection (TB)
3- CML 4- whooping cough (pertussis)
5- Infection mononucleosis (detect by pual –bunnel test)
*Monocytosis: increased in monocytic count
Causes:
1- Infection (TB) 2- typhoid fever. 3- Hodgkin's disease.
*Leucopenia: Decreased in WBCs number.
*Neutropenia: Decreased in neutrophil number.
Causes:
1- Drugs: cytotoxic drugs 2- Immune: SLE 3- X-Ray
4- Infections: a-Viral: hepatitis b- Bacterial: TB

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 ‫شادي الحربي‬ ‫ملخص علم الدم‬

*Lymphopenia: Decreased in lymphocytic number

Causes:
1- Infections: HIV 2- Drugs: cytotoxic drugs
3- Radiotherapy 4- Immune: SLE
5- Cushing's syndrome 6- Hodgkin's disease

*ESR(Erythrocyte Sedimentation Rate):

-Pipette used for it called Westergen's pipette.
-N.R: Male < 15 mm\hr Female < 20 mm\hr
*ESR Depends on:-
1- Plasma protein change. 2- Hb Concentration.
3- RBCs changes.
*ESR Very High (100 mm\hr )) the pathognomonic of :
1- Malignancy 2- autoimmune disease (SLE)
3- Active TB 4- Myeloma 5- Rheumatic Fever
* Diseases associated with normal ESR:
1- Polycythemia 2- viral diseases
3- Sickle cell anemia 4- congestive heart failure

Hemoglobin{Hb}:-
N.R Hb = 12 – 17 g/dl.
Hb F:- 2% in adult. Most found in children70%.
2 ά chains and 2 γ chains.
Hb A:- Most found adult 96 % .
2 ά chains and 2 β chains.
Hb A2: 2 %
2 ά chains and 2 δ chains.
Hb S: found in sickle cell anemia.
Hb H:
Hb C:
Hb E:

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Anemia
-It is reduction in the amount of circulating Hb, RBCs or both.
General Classification of Anemia:4
A-Microcytic anemia: (MCV less Than 80 FL)
1- Iron deficiency anemia. 2-Sidroblastic anemia. 3-Anemia of chronic disorder.
4-Lead poisoning. 5-Thalassemia.
B-Macrocytic: (MCV more than 100 FL)
1- Megaloblastic anemia (Pernicious anemia, Folic acid deficiency anemia).
2- Alcoholism.
C-Normocytic anemia: (MCV is Normal 80-100 FL)
-Aplastic anemia (AA).
D-Hemolytic anemia:
A-Congenital hemolytic:
1-Membrane defect: (Hereditary Spherocytosis, Hereditary eliptocytosis).
2-Enzyme defect: (G6PD deficiency).
3-Hb defect: (Sickle cell anemia, Hb pathies).
B-Autoimmune hemolytic anemia
-(AID (SLE), Blood group incompatibity).

A-Microcytic anemia:
-all are microcytic hypochromic in P.B.P except sidroplastic may be dimorphic picture
(normocytic normochromic or microcytic hypochromic).
-Hb, PCV, MCV, MCH are reduce in all and MCHC is normal (not affected) in all.
1-Iron deficiency anemia:
-It is the commonest anemia in the world.
-Iron deficiency develop slowly and symptoms only seen when Hb <8g/dl.
-The Iron store in liver in the form of Ferritin and Hemsiderin.
- Ferritin circulated in plasma but Hemsiderin cannot enter the circulation only in liver.
-Cause plumer –vinson syndrome and pencil cells shaped may target cells.
Diagnosis:
1- Microcytic Hypochromic (MCV< 80FL), (MCH<27pg).
2- S.Iron (N.R 60-190 ug) and S.Ferritin are decrease.
3-TIBC is increase. 4-Free erythrocyte protoporphyrin is increase.
5-Percentige saturation of transferrin is decrease. N.R 35-40%.
6- Erythropoiesis is normoblast . 7-Target cells.
8-Decreased or absent of hemosiderin stained by Prussian blue stain to give golden–
brown and blue granules.

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2-Sidroblastic anemia:
A-Hereditary Sidroblastic anemia: related with chromosomal abnormalities.
B-Acquired Sidroblastic anemia: due to chemical or drugs.
-The defect in protoporphyrin due to deficiency of delta amino lesulimik synthetase
enzyme which responsible for synthesis of protoporphyrin. Lead to decrease
protoporphyrin and increase S.iron.
*Sidrocyte: Mature RBCs containing non heam iron but cannot appear in PBP
because it removed by spleen.
*Sidroblast: Immature RBCs in B.M.
-Type I:
-Contain iron but cannot utilize for Hb synthesis. Hb is normally synthesis in this type.
-30-40% normally found in circulation.
- Type II:
- Contain iron but cannot utilize for Hb synthesis. Hb is normally synthesis and iron in
this type more than type I.
-Type III:
- Contain iron but cannot utilize for Hb synthesis.
-Hb is abnormal and iron form granules around the nucleus called ring sidroblast.
Diagnosis:
-The diagnosis is closely opposite to iron deficiency anemia except dimorphic picture
present only in sidroblstic anemia (normcytic normochromic or microcytic).

3-Anemia of chronic disorder:

-Characterized by decrease releasing of iron from macrophage to plasma and so to
erythroblast.
-Causes:
1-Chronic inflammatory disease such as T.B, rheumatic arthritis & SLE.
2-malignant disease such as carcinoma, lymphoma & sarcoma.
- Diagnosis:
1-S.iron decrease. 2-TIBC decrease.
3-S.firritin increase (Fe is present but cannot able to reach circulation).
4-Percentige saturation of transferring is Normal.
5-Free protoporphyrin Increase. 6-S.Transferrin decrease. #################

4-Lead poisoning:((‫التسمم بالرصاص‬####################

-Lead to hemolysis of RBCs due to poisoning and patients are usually death before
diagnosis in the lab.

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 ‫شادي الحربي‬ ‫ملخص علم الدم‬

5-Thalassemia:
-The defect of thalassemia occurs in globins gene.
1-Alpha thalassemia:
-The defect in alpha globins gene due to deletion of 1 or 2 or 3or 4 alpha globins gene.
-Deletion in one gene called Silent carrier.
-In 2 gene called alpha thalassemia trait: (RBCS Increase).
-In 3gene called Hb H disease or beta 4: (golve ball inclusion bodies).
-In 4 genes called hydrofetalis Hb or Hb Bar disease (4 gammas): the patient
is in compatible for live due of 4 gamma are high affinity to oxygen.
-The symptoms of this are depend on the number of gene deleted.
2-Beta thalassemia:
-The defect in beta globins gene.
A-Beta thalassemia major (Coolys anemia):
-Homozygous.due to total absence of beta globin gene production or the beta globins
gene are present but not functional.
B-Beta thalassemia minor:
-Heterozygous carrier and usually no clinical symptoms.
Diagnosis of Thalasemia:
1-All are Microcytic Hypochromic.
2-Sever anemia Hb 2-3g/dl in Coolys anemia.
3- Defective in Hb F (more than 70% or 90% in Hb electrophoresis test).
4- Absence or decrease of HbA.
5-Basophilic stippling. 6-nucleated red cells. 7-Target cells.
8- S.iron, S.ferritin, TIBC, Free protoporphyrin and percentage saturation of transferrin
are normal because the defect in globins gene not in heam.

B-Macrocytic: (MCV more than 100 FL)

1-Megaloblastic anemia
- Due to deficiency of Vitamin B12 or Folic Acid or both.
- Function of Vitamin B12 and Folic Acid:
-It is important in formation of DNA & RBCs.
A-Pernicious anemia:
-Causes due lack of Intrinsic factor or gastrictomy.
- I.F transport Vit B12 from stomach to the site of absorption (terminal Ilium).
-Transcoblamin 1(alpha globulin-not effect-granulocyte) or Transcoblamin 2(Beta
globulin-V.I) transport Vit B12 from terminal Ilium to B.M for uses or to site of
storage in liver.
-Cause Neurological symptoms. Howell-jolly bodies and Cabot ring in severe cases.
-Diagnosis by Schilling test.
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Vit B12:
-The daily requirement of vit B12 in the body is 2ug. N.R=3-5 mg.
-Any deficiency in vit B12 with normal I.F called megablastic anemia
- Vit B12 synthesis by M.O inside the body but cannot absorb by body.##########
-It is a disease of later life usually age of > 40 years.
- Vit B12 store mainly in liver called adenocylecobalomin.
- Vit B12 in plasma called methyl coblamin.
-Diphylobothrium latum parasite cause Vit B12 deficiency because it is taking up
from blood and cause megablastic anemia due to this.
B-Folic acid deficiency anemia:
-The daily requirement of folate in the body is 100-200 ug.
-Cause macrocytic anemia due to Folate deficiency.
-Absorption in duodenum and upper duodenum.
- Folate in plasma called methyl tetrahydroflate monoglutamate.in RBCs and liver
called methyl tetrahydroflate polyglutamate.
-The biochemical active residues of folic acid is dihydrofolate.
-Not cause Neurological symptoms and sore tongue.
Diagnosis of Megaloblastic anemia:
1- Macrocytic anemia (MCV more than 100FL)
2- Decreased Serum Level of Vit B12 or Folic Acid.
5-Pancytopenia +hyper segmented neutrophil. V.I
3- Horse shoe shaped cell. 4- Reduce red cell life span.
2- Alcoholism:
-Any liver disease cause round macrocytic anemia not megablastic.

C-Normocytic anemia: (MCV is Normal 80-100 FL)

Aplastic anemia(AA):
-It is pancytopenia due to Bone Marrow failure or decrease B.M production.
Causes:-
A- Congenital: fanconi's anemia defect in DNA.
B- Acquired: Idiopathic
1- Drugs (chloroamphinicol). 2- Chemical: Benzene 3- Radiation
4- Viruses: (hepatitis C, Epstein-Barr virus)
Diagnosis of aplastic anemia:
1-Normocytic normochromic or macrocytic 2-Pancytopenia.
4- marked increased of erythropoietin hormone level in serum and urine.
6-Neutrophil alkaline phosphates Increase. 5- Enlargement of spleen.

*Pure red cell aplasia: -Affect only the erythropoietin cells.

-patient with aplasia have anemia + reticuloculocytopenia with normal WBC and
platelet count.
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D-Hemolytic anemia:
Clinical feature of hemolytic anemia:
1- Normocytic normochromic except MCHC is normal or increase in Spherocytosis.
2- Reticulocytosis (Erythroid hyperplasia).
3-Splenomegally except in Sickle cell anemia is Atrophy. V.I
4- Haptoglobin decrease. (Alphaglycoprotin) react with free Hb.
5-Pollar and mild jaundice.
7- Fragmented cells + Schistocyte. V.I
8-Ostomatocyte and acanthocyte.
9-Hemoglobinemia: free Hb in circulation.
-Plasma hemopoxin react with free Hb converted to mess Hb then dissociated.
-Any inclusion body not appear in circulaton because it removed by spleen.

1-Hereditary Spherocytosis:
-The defect in cell membrane proteins (Spectrin) either:
Quantitative: due to decrease amount of cell membrane proteins (Spectrin).
Qualitative: the proteins (Spectrin) is present but not functional.
Any defect in cell membrane proteins cause release of membrane lipid and lead to
decrease surface area. (Spherocyte).
Diagnosis of Spherocytosis anemia:
1-Spherocyte lacks the central parel.
2-Osmatic fragility test increase. V.I
3-Acidified glycerol lysis time.

2-Hereditary Eliptocytosis:
-Same of spherocytosis but deferent in PBP appear as eliptocytosis.

3-G6PD deficiency anemia (Favism):

-It is sex linked disorders due to defect in enzyme of G6PD.
-Any sex linked disorders carrier in female and diseases in male.
-Type B is common type. ##########################################
-The defective gene is present on the chromosome X, mainly seen in males.
-Female heterozygous G6PD deficiency can resistant the malaria falciprum.
-G6PD deficiency leads to NAPPH deficiency lead to precipitation of Heinz body.
-Heinz body stained by supravital stain.
-Pyruvate Kinase deficiency: lead to G6PD deficiency must be homozygous.
-Special test is enzyme level for G6PD and collected in EDTA anticoagulant only. V.I.
FAVISM:
-It is acute hemolytic anemia result from G6PD deficiency, usually effect children and
combined with Hb urea.
-Causes see Heinz bodies in the peripheral red cells.

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3-Sickle cell anemia(Drepanocytes):

-Caused by substitution of one aa (valine) instead of glutamic acid at position No 6 in
the Beta chain of Hb resulting in abnormal HbS.
-Sickle cell causes atrophy of the spleen. Not cause spleenomegaly.
-Sickle cell trait in heterozygous.#####################################
-ESR reduced only in hemolytic anemia.
Tests For diagnosis of Sickle cell anemia:
1- Sickiling test.
2- Hb electrophoresis (Hb A, Hb S, Hb F).
3-Hb S and Hb- ---are migrated as the same strand in electrophoresis.##########
4-Hb solubility test +ve.######################################

5-Hb pathies:
-It is condition due to qualitative defect in globins polypeptide chain.
Classified into 3groups:
A-Due to structure variant of Hb:
1-Single aa substitution: eg: Hb S, Hb C, and Hb E, Hb D.###############
Hb C:
-Caused by substitution of one aa (Lysine) instead of glutamic acid at position No 6 in
the Beta chain of Hb resulting in abnormal Hb C.
Target cells are Prominent in Hb C. V.I-
-In soluble and less soluble than Hb A.
-Blood incubation with 3%Nacl in W.P at 37 C.
-Wet preparation
-Hb C, Hb E, Hb A2 at the same position in electrophoresis technique.
-Hb C trait no anemic, Target cell.
Hb E:
-Caused by substitution of one aa (Lysine) instead of glutamic acid at position No 26
in the Alpha chain of Hb resulting in abnormal Hb E.
Hb D:
-Caused by substitution of one aa (Protamine) instead of glutamic acid at position No
121 in the Alpha chain of Hb resulting in abnormal Hb D.
2-Multi aa substitutions:
Eg: Hb bar
B-Due to failure synthesis of Hb normally:
Eg: Beta talassemia
C- Due to failure to complete new switch from Hb F to Hb A.
*Hb pathies abnormality due to change in behavior of Hb may be due to aggregation,
insolubility, high O2 affinity and instability (release of aa lead to precipitation and
denaturation of globin in side RBCs and RBCs become abnormal).
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B-Autoimmune Hemolytic anemia:

-It is group of hemolytic anemia result from Ab direct against self antigens.
-eg: IgG, IgM less common, IgA and some bind complement.
-It classified according to Temp and according to etiology.
A-according to Temp in to worm Ab and cold Ab:
1-Worm Ab:
-IgG react at 37C. –Most are anti Rh antigen.
2-Cold Ab:
a-Cold hemoaglutination disease:
-It is IgM react in 4 C. –Most are anti i and anti I.
b-Paroxysmal cold Hb urea (PCH):
-IgG react in 4 C. –Anti D
B-According to etiology:
-Idiopathic or secondary.
Lab diagnosis of worm Ab AIH:
-Hb, PCV decrease. WBCs, Plt normal. MCV mild increase, MCH, MCHC normal.
-In PBP spherocytosis and reticulocytosis. –polychromasia.
-To differentiate between spherocytosis in AIH and in hereditary make DAT is +ve in
worm AIH anemia.
-Direct comb test is +ve in all hemolytic anemia.
-S.folate and cell folate are decrease.
Lab diagnosis of cold PCH:
-Donald Landsteiner Ab test:
-It is special test for Hb urea.
*Relation between erythroid hyperplasia and reduced M/E ratio:-
-M/E ratio is: ratio between number of cell of neutrophile series and the number of
erythroblast in the bone marrow. Normal range: 2:8
- Reduction of M/E ratio is taken as evidence of erythroid hyperplasia.

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 ‫شادي الحربي‬ ‫ملخص علم الدم‬

Leukemia
-It is malignant proliferation of hematopoietic cells characterized by abnormal
accumulation of WBCs in B.M and peripheral blood.
Causes of leukemia:
1-unknown causes. 2- Ionizing radiation.
3- Drugs and chemical. 4- Genetics. 5- Retrovirus.
6- Immune states.
Classification of leukemia:
A- Acute leukemia: (high malignancy).
- It is malignant proliferation of Immature hematopoietic cells (blast cells).
1-AML:
- Most common in adult. With Auer rods.
-have 8 subtypes from m0 to m7.
2-ALL:
-Most common in children.
-have 3 sub types L1, L2, L3.
B- Chronic leukemia: (Low malignancy).
-It is malignant proliferation of mature and immature hematopoietic cells.
1-CML:
-Most common in adult and diagnostic by Philadelphia (Ph)-chromosome.
- Philadelphia chromosome due to cross link between long arms of chromosome 9
with long arm of chromosome 22.
-Neutrophil alkaline phosphates are decrease.
2-CLL:
-It is accumulation of abnormal lymphocyte in B.M and peripheral blood.
-Smudge cell character in peripheral blood.

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 ‫شادي الحربي‬ ‫ملخص علم الدم‬

Leukomoid reaction:
-Leucocytosis characterized by presence of Immature cells and high neutrophil
alkaline phosphates.
-We can distinguish it from CML by:
1-Malignant tumor 2-Acue infection
2-Acute hemolysis. 3-Burns PP.
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HEMOSTASIS
-It is the mechanism by which the blood is kept in fluid state inside the blood vessels.
Function of homeostasis:
A-To stop blood bleeding by:
1-Blood vessels. 2-Platelets. 3-Coagulation factors.
B-To prevent thrombosis by:
1-Blood flow. 2-Platelets. 3-Inhibitors 4-Fibrinolytic system.
1-Blood vessels (B.V):
-Intima of B.V consists from endothelial (non thrombogenic layer) and sub endothelial.
-The sub endothelial layer contain:
1- -ve charge collagen which responsible for activation of Plt and C.F.
2-Elastic fiber 3-Protoglycan or glycoprotein.
Functions of endothelial layer:
1-produce angiotensin II (vasoconstriction).
2-produce Vonwillebrand factor(VWF) (50% carry factor VIII in the circulation and
50% enter the sub endothelial layer).
3-glycocaylx contain heparin sulphate which activating antithrombin III (strong
inhibiter).
4-produce prostacyclin which prevent Plt aggregation in normal condition.
5-contain receptor thrombomodulin which bind with thrombin to activate protein C
to inhibit factor Va ,VIIIa.
6-Thrombin activate endothelial to produce plasminogene.
7-Synthesis protein S which is cofactor for protein C.
2-Platelete (Plt):
-Discoid, non nucleated and formed in the B.M by megakaryocytic.
-Life span is 10 days and N.R 150000-450000/L.
The cytoplasm of Plt contains 3 types of granules:
1-Dense granules: contain ADP, ATP, and Serotonin when release in circulation act
as agonist.
2-Alpha granules: produce PDGF to act on B.M to increase production of Plts.
3-Lysosomal granules: produce lysosomal enzyme which help in destruction of any
invading M.O and hence the Immuno response.
Functions of Plt: formation of haemostatic plug at the site of damage by:
1-Adhesion 2-Release reaction 3-aggregation
4-procoagulant activity: -contain activation for factor XI.
-Source of tissue thromboplastin and phospholipids.
-Plt contains TXA2.(agonist)(short life span 1min).

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3-Coagulation factors (C.F):

-It is plasma protein circulate in active form called pro-enzyme.
-All C.F produce by liver except VIII are produce by endothelial cells.
Classification of C.F based on functional and biochemical properties into 4 groups:
1-Vit K dependant C.F:
-Factor II, VII, IX, X, Protein C and S.
-Warfarin is oral anticoagulant which stop the activity of Vit K (PIVKA state) Vit K
is present but not functional.
2-Contact factors:
- Factor I, XI, XII, Prekallekerin.
3-Thrombin sensitive factors:
-Factor I, V, XIII.
4-Labile factor:
-Factor VIII, V.
- Factor VIII circulates in combination with VWF.
Activation of C.F by 3 pathways:
1-Internsic pathway:
-Contain –ve charge collagens fiber, XII, XI, IX (VIIIa+Ca+phospholipid) called
protease enzyme, X.
2-Externsic pathway:
- Contain Tissue thromboplastin (damage tissue+plat), VII, X.
3-Common pathway:
-Contain X, Prothrombinase enzyme(Va +Ca+ phospholipids), prothrombin II,
thrombin IIa, fibrinogen I, Fibrin Ia(monomer).
-Thrombin IIa activate V to Va and VIII to VIIIa and XIII to XIIIa. And also
activate protein C, S. system inhibitor.
- Ia(monomers) polymerization by the action of XIIIa to Ia(polymers).
-Plts has receptor called fibrinogen receptor attach to Ia polymers to form secondary
haemostatic plug.
Type of Inhibiters:
1-blood flow. 2-Serine protease 3-fibrinolytic system.
Test used to asses haemostatic mechanism:
A-Bleeding time(BT):
-This test depends on platelets and B.V (capillary fragility).
1- IV method: normal 1- 4 min with IVY method.
2-Dukes method: normal 4-6 min.
3-Standraized template method: normal 2-6 min.
B-Clotting Time(CT):
-Screening test for all C.F I – XIII except VII.
-Used whole blood clotting time with water path at 37. Normal time = 3 – 9 min.

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C-Plasma clotting time:

-Include: PT, PTT, TT.
1- Prothrombin time (PT):
- Screening test for Extrinsic and Common pathway.
-Consist of 2 factors: a- III Tissue thrombo blast b- VII
- It is prolonged when deficiency of ( I , II , V , VII , X ).
- Sensitive special to deficiency of (V, VII, X).
- We measure PT by adding thromboplastin, Calcium to activate extrinsic path way.
Prothrombin ratio % = PT of test
PT of control
-International Normalized Ratio (INR) = PT of test
Or Sensitivity of reagent (IST) PT of control
-Prolong: defect in liver or Vit K deficiency or oral anticoagulant.
- This test used for control or anticoagulant treatment (warfarin).
-Normal time = 10– 15 sec
2- Partial thromboplastin time (PTT,APTT,KCCT):
- Screening test for Intrinsic and Common pathway.
-Consist of 6 factor (XII(Prekallerin, HmwK), XI, IX, X, II, I)V,.
-It is prolonged when there is deficiency of factor (VIII-IX).
-When VIII deficiency it causes:
a- hemophilia A. b- Von willebrand disease.
-when IX deficiency it causes:
a-hemophilia B (Christmas disease)
We measure PTT by adding:
a- kaolin (KCCT) b- phospholipids c - Calcium.
- Normal time = 30 – 40 sec.
-This test used for control or anticoagulant treatment (heparin).
3- Thrombin time (TT):
- Screening test for fibrinogen (detect defect or deficiency of fibrinogen).
-It is prolonged when there is deficiency of fibrinogen and presence of heparin.
- Conversion prothrombin to fibrinogen by adds thrombin.
-Normal time = 10 – 20 sec.
D- Clot solubility test:
-Specific test for XIII.
-Use 1% of monochloroactic acid.
-Fibrin polymers: clot remains for 2hrs without dissolving.
-Fibrin monomers: clot dissolved < 2hrs (XIII deficiency).

*Antithrombin III: VI
-It is mainly inhibitor of thrombin and also factors IXa, Xa, XIa.
-It is potentiated by heparin.
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There is 6 factors should be known: V.I

1-VIII anti-hemophilic. 2- IX Christmas.
3- II prothrombin. 4- IIa thrombin.
5- I fibrinogen. 6- Ia fibrin.
Protein C and protein S:
-Protein C is protease and degrades Va , VIIIa.
-Protein S is potentiated and effect of activated protein C.
Some convert of factor: VI
-Plasminogen converts to plasmin by: a- kallikarin(very strong). b- Urokinase. c-
Streptokinase.
-Fibrin converts to fibrin degradation product by plasmin.
-Fibrinogen to fibrin by thrombin.
Anticoagulant drugs: VI
-Most common 2 drug heparin and warfarin.
1-Heparin:
-It is acidic mucopolysaccaride and does not cross placenta.
-It is potentiates action of anti thrombin III.
-Overdose of heparin cause hemorrhage managed by stopping heparin by giving
protamine sulphate.
2-Warfarin:
-It is Vit K antagonist and cross placenta.
-Cause hemorrhage managed by stopping warfarin and infusing fresh frozen plasma
and give Vit K.

Disorder of haemostatic mechanism:

-Due to bleeding tendency or thrombosis.
-In bleeding tendency the defect in primary H-disorder (B.V, Plts) or secondary
H-disorder (C.F).
-In thrombosis the defect in Inhibitor or fibrinolytic system.
Primary H-disorder: divided in to:
A- Disorder of vascular H:
-Mainly congenital due to elastic fiber deficiency or acquired due to collagen deficiency
resulting of increase vascular permeability. Diagnosis by clinical no specific test.
B-Platelets Disorder: either:
1-Numerical disorder:
-If the Plts < 50.000 cell/cum the bleeding occur.
-If the Plts < 10.000 cell/cum the bleeding occur spontaneously.
2-Funtional disorder: either:
a- Congenital: due to chromosomal abnormalities and divided into:
Receptor defect, enzyme defect, granules defect.

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b-Acquired: such as: Aspirin, heparin, multiple myeloma, myelofibrosis.

-Aspirin Inhibit cyclo oxygenase enzyme lead to decrease Thromboxane A2(agonist).
-Test used for primary H-disorder called Hiss test or tourniquet test: N.R < 20 dotes
Secondary H-disorder (C.F): either:
A-Congenital disorder:
-Always deficiency in simple C.F such as:
1-Haemophilia. 2-VWF disease. 3-Fibrinogen disorder.
B-Acquired disorder:
-Deficiency in multiple C.F such as:
1-DIC 2-Liver disease 3-Vit K deficiency.

Haemophilia A: Congenital deficiency of VIII called Classical hemophilia most

sever in Royal disease.
Haemophilia B: Congenital deficiency of IX called Christmas disease.
Haemophilia C: Congenital deficiency of XI.
-Hemophilia A and B are inherited as sex linked recessive.
-Male ---------hemophilia 50%normal 50%heamophilic
-Female-------carrier 50%normal 50%carrier (daughter)
-Para hemophilia deficiency in factor V.

blood bank and

serology
.Forward grouping: detecting ABO grouping by using the cells (RBCs) of the patient
.Reverse grouping: detecting ABO grouping using the serum of the patient

the color of anti-sera A == blue -

the color of anti-sera B == yellow -
the color of anti-sera D == gray -

.Antigen: substance capable of inducing specific immune response

.Blood group Ag is attached to cell membrane-
Antibody Ab (Immunoglobulin(IG): are the proteins that synthesized by
.B-lymphocyte
:Class of antibody-
.IgG.(lowest M.W) 2- IgM(highest M.W) 3- IgA. 4- IgD. 5- IgE-1
:ABO blood group system
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.The most blood group system-

.Essential for prevent of hemolytic transfusion reaction -

GROUP Antigen (AG) Antibody

(AB)
A A B
B B A
AB AB ------
O ------- AB

Recipient ABO Compatible donor blood

group
A A–O
B B–O
AB AB – A –B –O
O O
.Universal donor: O *
.Recipient donor: AB *

.Rhesus group: contain six Ags are: C- D – E –c –d – e*

.antigen D: is most antigens in this group-
the presence or absence of D-antigen determine if Rh positive or Rh negative-
.Fisher– race terminology is used to describe Rh antigen -
.If D antigen found Rh is positive (85%) -
.If D antigen not found Rh is negative (15%) -
.Du antigen is weak type of D antigen -

:Other blood group system*

kell blood group system : kell Abs are IgG, they can cause hemolytic disease for -1
. newborn
duffy blood group system: duffy Abs are IgG , they can cause hemolytic disease for-2
. newborn and detect malaria parasites
kidd blood group system: Kidd Abs are IgG , they can cause hemolytic disease for -3
. newborn

. HLA antigen: are controlled by the Chromosome 6 *

.Anti I :- it is react with all red cell except cord cells *

.Anti i :- it is react with cord cell*
.That means we can differentiate between anti I & anti i by cord cells -
-:Antiglobulin test (AHG)*
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.Antibody is gamma globulin -

.Most incomplete antibodies are IgG -
.AHG serum contains Anti-IGg and Anti C3-
.AHG serum used by most labs for pre-transfusion IAT Anti-IgG -
.when doing DAT to detect in vivo sensitization with IgG or C3-
-Anti-human Immunoglubin:-
1-coombs test. 2- anti-Ab = anti-antibody. 3-anti-IgG.
:Direct and indirect antiglobulin test (coomb's test) *
Binding of IgM to red cell is usually cause agglutination and IgG bind to red cell is -
.not cause agglutination
.IgG can be detect by antiglobulin test (coomb's test)-
:A-Direct antiglobulin test (DAT)
.Used to detect sensitized RBCs in patient blood-
:Application of DAT
.Hemolytic disease of newborn -1
Autoimmunohemolytic anemia -2
Drug induced hemolytic anemia -3
.Transfusion reaction -4
:B-Indirect antiglobulin test (IAT)
.Used to detect sensitized antibody in patient serum-
.the RBCs are washing several times to remove unbounded proteins (IGg)-
-:Application of IAT
.Cross matching -1
.Antibody Screening -2
.Phenotyping -3
-:Cross matching*
-:Types of cross matching
Major cross matching: To determine compatibility between Donor red cells -1
.and patient serum (plasma)
Minor cross matching: To determine compatibility between Donor serum and -2
.patient red cells
-:Common causes of false positive in both IAT and DAT *
.Cold agglutination -1
.Rouleaux -2
.Reagent contamination -3
.Antibody low incidence antigen -4
.Human errors -5
-:Common causes of false negative in both IAT and DAT *
.Failure to add antiglobulin reagent. (Anti Sera) -1
.Add wrong antiglobulin reagent (Anti Sera) -2
-:Hemolytic disease of newborn ( HDN) *
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.Can develop when mother Rh negative and Fetus Rh positive during pregnancy -
Mother Rh negative administrate Rh immunoglobulin ( RhIG) to prevent hemolytic -
.disease in newborn

:Selection of blood donor in KSA*

:blood donor must meet the following criteria-
.age: at least 17 years old-1
.weight: at least 50 kg -2
.Hb: 13-18g /dl -3
.three month from last donation -4
.Blood pressure: 120/70 -5
Body temperature: 37 C -6
Pulse rate: 70/min -7
:Adverse donor reaction *
.syncope (fainting): the most common donor adverse reaction-1
.Nausea and vomiting -2
.hematoma-3
:Serological test for blood donor*
HIV 1&2 2- HBV-1
HCV 4-HTLV-3
.Syphilis. 6- Malaria-5
:A-HIV (human immunodeficiency viruses)
the etiologic agent of AIDS-
:Serological test of AIDS
ELISA-1
.Western blot (WB) is confirmatory test -2
:B-HBV (hepatitis B)
.Serological test for HBV by ELIZA-
:C-HCV (hepatitis C)
Serological test
.ELIZA. 2-RIBA (confirmatory test) -1
:D-SYPHILIS
.is complex sexually transmitted disease-
:Serological test-
.RPR (rapid plasma region)-1
.VDRL -2
:E-HTLV (human T-cell lymphotropic virus)
:Serological test
1- Western blot (WB) is confirmatory test ELISA-2
:F-MALARIA
.Diagnosis by THICK FILM-
:Transfusion reaction screening of blood *

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:Transfusion screening test -

.Patient blood group & antibody detection -1
.Donor blood group -2
.Screening of donor blood for infectious disease -3
.Cross matching -4
:Anticoagulant and Preservative *
:Anticoagulant used for blood donor bags are-

Preservative Temp Expiration

citrate – phosphate –dextrose (CPD)-1 C 1-6 21
citrate –phosphate–dextrose- dextrose -2 C 1-6 21
(CP2D)
citrate –phosphate –dextrose-adenine -3 C 1-6 35
(CPDA-1)
.CPDA-1 is most common anticoagulant used in blood bank-
- The storage time of saline adenine glucose mannitol (SAGM) is 42 days
.Citrate: prevent coagulation-
.Phosphate: act as buffer to decrease PH-
.Dextrose: used for ATP generation -
.Adenine: substrate for the red cell to synthesize of ATP -
:Storage of blood component*
Component Blood Expiration
Whole blood C 1-6
RBC 1 years (-80C)
Platelet days(20-24 C) 5
Fresh Frozen Plasma FFP years (-18 C) 1
Cryoprecipitate years (-18 C) 1
:Red Cell Component*
.Whole blood -1
:A-RBC (packed red cell)
.RBCs are unit of whole blood with most of the plasma removed-
RBC: is the component of choice symptomatic anemia also used for hemolytic -
.newborn exchange transfusion
.RBC can be store by frozen for years by adding: -Glycerol -
:B-WBCs
-:can be reduced in the whole blood bag by several methods
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.Centrifugation. 2- Saline washing. 3- Freezing. 4- Filtration -1

.Filtration is the best method choice today in blood bank-
:C-Plasma Components
:Fresh Frozen Plasma FFP -1
-:Used to treatment for patient associated with-
.a- Clotting factor deficiency
.b- Disseminated intravascular coagulation (DIC)
.c-Vit K deficiency
:Cryoprecipitate-2
It is cold- Insoluble portion of plasma that precipitates when FFP is thawed between-
.1-6C
Used to control bleeding associated with fibrinogen deficiency or factor XIII -
.deficiency
-:Cryoprecipitate also used for treatmentment
.hemophilia A. 2- Von willebrand disease-1
:D-Platelets
Used for treatment of bleeding in patient with thrombocytopenia or platelet -
.dysfunction
.Platelets are extremely sensitive to temperature and PH changes-

*:Store Temprature for blood components

Components Temperature
Whole blood C 45 days 1-6
Red blood cell (RBC) C 1-6
Fresh frozen plasma (FFP) years (-18 C) 1
Platelets days(20-24 C) 5
Granulocytes C 20-24
Cryoprecipitate years (-18 C) 1

*Thrombocytosis:
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-It is increased platelets count.

Types: 1-Primary (essential). 2-Secondary
-------------------------------------------------------------------------------------------------

*Thrombocytopenia:
-decreased in platelets count
Causes:
1-Aplastic Anemia. 2-Viral: CMV
3- Idiopathic thrombocytopenic purpura ( ITP).
4-Malignancy or leukemia 5-Drugs

*multiple myeloma:
-Disease arising from malignant transformation of B cell.
-Most common symptoms present is bone – pain.
Diagnosis of multiple myeloma:
-Normocytic+ Normochromic: (common)
1-Bence –jonec protein present (important)
2-thrombocytopenia + neutropenia 3-increased basophile.
4-Increased red cell 5-ESR high 6-plasma cell high
7-monoclonal Ig in serum, monoclonal light chain in urine.
*Malignant lymphoma:
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-Tumor of lymphoid tissue.

*Hodgkin's disease:
-This diseases affect the lymph tissues.
-Large binucleate or multinucleate cell called Reed –Sternberg cells.
-Affected lymph node contains Epstein Barr virus genome.
Diagnosis:
1-normocytic + normochromic anemia
2-high ESR 3- neutrophilia 4-eosinphillia
5-lymphopenia
*NON Hodgkin's disease:
-The etiology is unknown
-This disease associated with patient they have acquired immune deficiency (AIDS).

*Systemic Lupus Erythematosis (SLE):

It is Autoimmune Disease affecingt many Organs .
*Tests of SLE:
- by Detection of anti-body (Abs):
1- antinuclear Ab (ANA): But not specific test for SLE.
2- Anti-Double Stranded- DNA Ab (Ads DNA): it is specific test for SLE
* Diagnosis of SLE:
- Blood Analysis: 1- pancytopenia 2- high ESR
-Urine analysis: 1- proteinuria. 2-hematuria. 3-RBCs &WBCs Casts.

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