Differential diagnosis of Anemia: 1. A complete blood count, CBC RBC count Hematocrit (Hct) or packed cell volume Hemoglobin determination RBC.
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<ul><li> Slide 1 </li> <li> Differential diagnosis of Anemia: 1. A complete blood count, CBC RBC count Hematocrit (Hct) or packed cell volume Hemoglobin determination RBC indices calculation Reticulocyte count 2. Blood smear examination to evaluate: Poikilocytosis. Leukocytes or Platelets abnormalities. </li> <li> Slide 2 </li> <li> 3. A bone marrow smear and biopsy to observe: Maturation of RBC and WBC series Presence of megakaryocytes Ratio of myeloid to erythroid series. Presence or absence of granulomas or tumor cells 4.Hemoglobin electrophoresis 5. Antiglobulin test: (Coombs test). 4. Osmotic fragility test </li> <li> Slide 3 </li> <li> Red blood cell indices RBCs Indices: They are part of the complete blood count (CBC) test that provide information about the hemoglobin content and size of red blood. They are used in diagnosis of anemia and its causes. The indices include: Mean corpuscular volume (MCV): is the average size of a red blood cell and is calculated by dividing the hematocrit (Hct) by the red blood cell count. MCV = Hct / RBC Normal range: (78-98) fL (femto- is 10 -15 ) </li> <li> Slide 4 </li> <li> Mean corpuscular hemoglobin (MCH): is the average amount of hemoglobin (Hb) per red blood cell and is calculated by dividing the hemoglobin by the red blood cell count. MCH = Hb / RBC Normal range: 27-32 pg/cell (pico- is 10 -12 ) Mean corpuscular hemoglobin concentration (MCHC): is the average concentration of hemoglobin per Packed red blood cells and is calculated by dividing the hemoglobin by the hematocrit. MCHC = Hb / Hct Normal range: 30-35 g/dL (deci is 10 -1 ) Red blood cell indices </li> <li> Slide 5 </li> <li> Morphological Classification of Anemia Based on RBC morphology Anemia is divided into three groups mainly on the basis of the MCV, and MCH (RBC indices): 1.Normocytic Normochromic anemia: (normal red cell indices) Blood loss anemia (Acute bleeding) Hemolytic anemia (except thalassaemia) Aplastic anemia Pure red cell aplasia. </li> <li> Slide 6 </li> <li> 2. Microcytic hypochromic anemia: ( low red cell indices) Iron deficiency anemia Sideroblastic anemia Thalassemia Chronic diseases 3. Macrocytic Normochromic ( high MCV and MCH, normal MCHC) Megaloblastic anemia (Vit. B12 deficiency & Folic acid deficiency). Liver disease Post splenectomy </li> <li> Slide 7 </li> <li> Morphological Classification of Anemia </li> <li> Slide 8 </li> <li> 1- Microcytic- Hypochromic Anemia. </li> <li> Slide 9 </li> <li> Slide 10 </li> <li> A- Iron Deficiency Anemia (IDA) : Is a condition in which the total body iron content is decreased below a normal level This results in a reduced red blood cell and hemoglobin production More than half of all anemia are due to iron deficiency. Clinical Picture: Symptoms: fatigue, dizziness, headache Signs eg. Pallor Tongue atrophy/ glossitis - raw and sore Angular cheilosis (Stomatitis) Spoon shaped nails (koilonychia), brittle nails and hair. </li> <li> Slide 11 </li> <li> Clinical Picture of IDA:, </li> <li> Slide 12 </li> <li> Lab. Investigations of IDA: a) CBC: Lab findings Low RBC, Hb, Hct Low MCV, MCH, MCHC Normal WBC and PLT RBC morphology Hypochromia Microcytosis Anisocytosis Poikilocytosis Pencil cells. Target cells </li> <li> Slide 13 </li> <li> b) Bone marrow iron (Tissue iron): - Erythroid hyperplasia. Tissue biopsy of bone marrow Prussian blue stain Type of iron is hemosiderin Absence of iron stores in BM. c) Plasma Iron parameters: Low serum iron, Low serum ferritin Iron deficiency </li> <li> Slide 14 </li> <li> B- Thalassemia: Inherited decrease in alpha or beta globin chain synthesis associated with Hb A quantitative defect All have microcytic/hypochromic RBCs and target cells Genetic mutations classified by: beta chains = beta thalassemiaGreek/Italian alpha chains = alpha thalassemiaAsian </li> <li> Slide 15 </li> <li> Slide 16 </li> <li> Classification of Thalassemia: These genetic disorders can be classified according to the severity. Severity ranges from lethal, to severe transfusion-dependency, to moderate, to no clinical abnormalities. Severity depends on the number and type of abnormal globin genes inherited. Clinically, Thalassemia are divided into: 1.Major severe anemia; no (or ) chains are produced, so cannot make normal hemoglobin. 2.Intermedia moderate anemia with splenomegaly & iron overload. 3.Minor/trait mild anemia; slight decrease in normal hemoglobin types made. </li> <li> Slide 17 </li> <li> Beta Thalassemia Major (Homozygous) Both beta genes are abnormal:(Chr. 11) Marked decrease/absence of beta chains leads to Hb F production ( 2 2 ) Hb A will not be produced. Rigid RBCs with Heinz bodies destroyed in bone marrow and blood (ineffective erythropoiesis). </li> <li> Slide 18 </li> <li> Beta Thalassemia Minor (Heterozygous) One abnormal beta gene allele: (Both Hb A and Hb F will be produced). Slight decreased rate of beta chain production Blood picture can look similar to iron deficiency </li> <li> Slide 19 </li> <li> Alpha Thal Major (Homozygous) Deletion of all 4 alpha genes (Ch. 16) results in complete absence of alpha chain production. No normal hemoglobin; Barts Hb formation (four chains Hb); Barts Hydrops Fetalis. Die of hypoxia. Barts Hb has a High affinity for oxygen. </li> <li> Slide 20 </li> <li> Alpha Thalassemia Intermedia = Hb H Disease: Three alpha genes deleted Moderate decrease in alpha chains leads to beta chain excess unstable Hb H (Excess 4 ). Moderate anemia. Formation of Barts Hb (4 ). Hepatosplenomegaly and H. bodies. Target cells </li> <li> Slide 21 </li> <li> Alpha Thalassaemia Minor (Heterozygous) One or two alpha genes deleted (group) Slight decrease in alpha chain production Mild or no anemia, few target cells Essentially normal electrophoresis; may undiagnosed </li> <li> Slide 22 </li> <li> Lab. Investigations of Thalassemia: Laboratory findings: Hb , MCV , MCH , MCHC RBCs : Hypochromic, Microcytic RBCs. Target cells, nucleated red cells Anisocytosis Poikilocytosis Basophilic stippling RBC inclusions Platelets: Normal WBCs: Normal Plasma: - iron - Normal or ferritin. </li> <li> Slide 23 </li> <li> 2. Macrocytic Normocytic Anemia: A. Megaloblastic anemia: Vitamin B 12 deficiency Folate deficiency Abnormal metabolism of folate and Vit. B 12 B. Non- Megaloblastic anemia: Liver disease Alcoholism Neonatal macrocytosis Stress erythropoiesis. </li> <li> Slide 24 </li> <li> A Mild to severe anemia, Increased MCV & MCH, normal MCHC Low RBC, Hb, WBC and PLT counts (fragile cells) due to ineffective hematopoiesis. Macrocytic ovalocytes and teardrops; Neutrophils show nuclear lobulations (hypersegmentations > 6 nuclei) Giant platelets & megakaryocytes fragments Lab Findings of Megaloblastic Anemia </li> <li> Slide 25 </li> <li> . </li> <li> Slide 26 </li> <li> 3. Normocytic Normochromic Anemia Is a condition in which the size & Hb content of RBCs is normal but the number of RBCs is decreased. It includes: Aplastic anemia due to BM failure Blood loss anemia Hemolytic anemia A.Aplastic Anemia Condition of blood pancytopenia caused by bone marrow failuredecreased production of all cell lines and replacement of marrow with fat. Due to damaged stem cells, damaged bone marrow environment or suppression. </li> <li> Slide 27 </li> <li> Lab diagnosis of Aplastic Anemia: 1- Peripheral blood: Normochromic Normocytic RBCs (normal MCV & MCH) Pancytopenia No abnormal cells 2- Bone marrow: Hypocellular BM with increased: -Fat spaces. -Lymph cells -Plasma cells -Macrophages -Mast cells </li> <li> Slide 28 </li> <li> B. Hemolytic anemia (H.A): Result from an increase in the rate of pre mature red cell destruction. It leads to Erythropoietic hyperplasia BM produces red cells 6 to 8 times the normal rate. Marked reticulocytosis. Two main mechanisms for RBC destruction in HA Intravascular hemolysis: in the circulation Extravascular hemolysis: in RE system (Reticuloendothelial system). </li> <li> Slide 29 </li> <li> Lab features of extravascular haemolysis: Increased RBC break down Serum bilirubin increase Stool stercobilinogen increase Urine urobilinogen increase Lab features of intravascular haemolysis: Hemoglobinemia and hemoglobinuria Hemosiderinuria Reduced/absent Serum haptoglobin. </li> <li> Slide 30 </li> <li> . </li> <li> Slide 31 </li> <li> Types of Hemolytic anemia: 1. Hereditary hemolytic anemia Result of intrinsic red cell defects: Genetic defects: 1- Genetic mutant allele result in RBCs Membrane structural proteins defect ( Defective spectrin tetramers). A-Hereditary Spherocytosis: (Osmotic fragility test) B- Elliptocytosis: Cigar shaped RBCs. </li> <li> Slide 32 </li> <li> N 2-X-linked genetic mutant allele results in Metabolic defect: Example: G 6 PD deficiency. Denatured globin due to oxidants (free radicals) action; Heinz bodies formation; Removed by RES Macrophage; characteristic bite cells. </li> <li> Slide 33 </li> <li> N 3-Inherited defect in hemoglobin gene (Beta-globin gene; Chr. 11): Example: Sickle cell anemia Valine substituted for glutamic acid at 6 th of -globin; production of abnormal hemoglobin ; Hb S. 1-Hemoglobin S Disease: HbSS: Two sickle cell genes inherited (both beta chains are abnormal). Symptomatic after 6 months of age. Laboratory findings: -CBC: Severe anemia and target cells. -Sickling test: Positive: 2% sodium bisulfite induce solution turbidity. -Hb electrophoresis: No Hb A, >80% Hb S, Hb F. HGB S Disease (Hgb SS) Sickle cell Target cell </li> <li> Slide 34 </li> <li> n Hemoglobin S trait /Sickle cell trait /Hb SA: No anemia or sickle cells. Target cells present. ~60% Hgb A, ~40% Hgb S </li> <li> Slide 35 </li> <li> Pattern of Sickle Hb Inheritance: N </li> <li> Slide 36 </li> <li> Hb Electrophoresis: Pattern of hemoglobin electrophoresis from several different individuals. Lanes 1 and 5 are hemoglobin standards. Lane 2 is a normal adult. Lane 3 is a normal neonate. Lane 4 is a homozygous HbS individual. Lanes 6 and 8 are heterozygous sickle individuals. Lane 7 is a SC disease individual. </li> <li> Slide 37 </li> <li> N 2. Acquired hemolytic anemia: A variety of acquired conditions result in: shortened survival of previously normal red cells. Result of extrinsic causes: 1-Immune Hemolytic anemia IHA; Warm IHA, Cold IHA (detected by Coombs test; Antiglobulin test). 2-Drug-induced HA. 3-Infection associated: Malaria. </li> </ul>
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