Group Members Aqsa Abbasi Ayesha Sajid Azka Khan Midhat Mustafa Sadia Arshad Fatima Riaz

1. Routine Hematology Tests Group Members Aqsa Abbasi Ayesha Sajid Azka Khan Midhat Mustafa SadiaArshad Fatima Riaz RidaRehman

2. What is Hematology • Full Blood Count (FBC) or Full Blood Examination (FBE) • The study of blood, the blood-forming organs, and blood diseases. • The study of etiology, diagnosis, treatment, prognosis, and prevention of blood diseases that affect the production of blood and its components. • These results are used in conjunction with the history, physical exam and other laboratory findings. • Qualitative and Quantitative Tests • Hematology tests helps in: • treating bleeding disorders • treating hematological malignacies • Screen, diagnose and measure the severity of disease • in the science of blood transfusion and the work of a blood bank • in bone marrow and stem cell transplantation

3. Routine Hematolgy Tests Complete Blood Cell Count • A complete blood count, often referred to as a CBC, is a common blood test. • A CBC provides detailed information about three types of cells in the blood. • Alexander Vastem is the first person to use the complete blood count for clinical purposes. • A CBC is routinely performed during annual physical examinations

4. Blood cell count principles

5. Platelet Count • A platelet count is a test to measure how many platelets you have in your blood. • They are smaller than red or white blood cells. • Normal Range: 150,000 - 400,000 platelets per microliter • Thrombocytopenia maybe due to • Cancer chemotherapy • Hemolytic anemia • Idiopathic thrombocytopenic purpura Leukemia • Massive blood transfusion • Thombotic thrombocytopenic purpura (TTP) • Thrombocytosis may be due to: • Chronicmyelogenousleukemia  (CML) • Polycythemiavera • Primary thrombocythemia • Recent spleen removal

6. CBC (RBCs, WBCs) Ayesha Sajid

7. RED BLOOD CELL TEST HEMOGLOBIN • Total amount of oxygen carrying protein inside RBCs • RANGE • Male 14-18 Gm/100 ml • Female 12-16.4 Gm/100 ml Number of RBCs per volume of whole blood RANGE Male 4.5-6.4 Mil/c mm Female 4.0-5.4 Mil/ c mm HEMATOCRIT • Percentage of blood volume made up of RBCs • RANGE • Male  42%-54% • Female 38%-46%

8. RED BLOOD CELL INDICES • TYPES OF ANEMIA • Microcytic anemia: small RBCs • Normocytic anemia: Normal red blood cell • Macrocytic anemia: larger than normal red blood cell

9. TOO MUCH VS TOO LOW

10. WHITE BLOOD CELLS Measures total white blood cellcomponent of whole blood RANGE Males: 5,000 to 10,000 per microliter (µL) of blood Females: 4,500 to 11,000 per microliter (µL) of blood PRINCIPLE Whole blood is diluted 1 in 20 in an acid reagent and white blood cells are counted

11. DIFFERENTIAL WBCs COUNT

12. LEUKEMIA

13. RETICULOCYTE COUNT Azka Khan

14. Reticulocyte Count Reticulocytes • immature erythrocytes • slightly larger than totally mature erythrocytes • Non-nucleated. • about 1% of the red cells in the human body • develop and mature in the red bone marrow and then circulate for about a day in the blood stream before developing into mature red blood cells. • Spend 2 to 3 days in bone marrow. • 1 day in peripheral blood before developing into mature erythrocytes. • Do not have completed production of hemoglobin and consequently contain residual hemoglobin synthetic machinery, mRNA and rRNA, commonly referred to as “reticulin’’. • They are called reticulocytes because of a reticular (mesh-like) network of ribosomal RNAthat becomes visible under a microscope with certain stains such as new methylene blue.

15. Reticulocytes Count A reticulocyte count measures the percentage of reticulocytes in the blood and assess the erythropoietic activity of the bone marrow. Why there is a need to perform • As a follow up to abnormal results on a CBC, RBC count, hemoglobin or hematocrit, to help determine the cause • To determine if the bone marrow is functioning properly and responding adequately to the body's need for red blood cells (RBCs) • To help detect and distinguish between different types of anemia • To monitor response to treatment, such as that for iron-deficiency anemia • To monitor bone marrow function following treatments such as chemotherapy • To monitor function following a bone marrow transplant.

16. High values A high reticulocyte count may mean more red blood cells are being made by the bone marrow. This can occur after a lot of bleeding, a move to a high altitude, or certain types of anemia. These conditions cause red blood cells to break down (hemolysis). The reticulocyte count rises after the treatment for pernicious anemia, iron deficiency anemia, or folic acid deficienc anemia starts working. Low values A low reticulocyte count may mean fewer red blood cells are being made by the bone marrow. This can be caused by aplastic anemia or other types of anemia, such as iron deficiency anemia. A low reticulocyte count can also be caused by exposure to radiation, a long-term (chronic) infection, or by certain medicines that damage the bone marrow.

17. PRINCIPLE • Whole blood anticougulated with EDTA, is stained with supra-vital dyes (stain living cells) such as methylene blue. • As reticulocytesare immature cells containing remnant cytoplasmic ribonucleic acid (RNA) and organelles such as mitochondria and ribosomes. • Reticulocytes are visualized by staining with supra-vital dyes that precipitate the RBC and organelles. This stain causes the ribosomal and residual RNA to coprecipitate with the few remaining mitochondria and ferritin masses in living young erythrocytes to form microscopically visible dark-blue clusters and filaments (reticulum). • less than 12 hours old is required. Normal range of reticulocyte

18. Prothrombin Time Evaluating Blood Clotting Efficiency

19. Physiology- The Story BehindHemostasis – Prothrombin’s Role in Coagulation

20. Significance – Why the test is performed?

21. Principle: How the test is performed?

22. Reference Ranges Source of Tissue Factor e.g. human, rabbit etc  - The exact technique used e.g. manual or automated • Each laboratory should establish its own normal range but in general the prothrombin time for a normal plasma sample, lies between 12-16 seconds   Method of end-point determination e.g. optical or mechanical

23. Atypical Results & Possible Reasons • Foods and substances that may alter PT • Alcohol • Oral contraceptives • Hormone replacement therapy (HRT) • Vitamin K excess – nutritional supplements and foods like broccoli, chickpeas, kale, liver (offal) and soybeans.

24. RIDA REHMAN IRON PROFILE

25. INTRODUCTION & SIGNIFICANCE • five different types of tests • Serum iron level test (SI) • total iron-binding capacity (TIBC) test • Ferritin tests • transferrin test • Transferrin saturation test • SIGNIFICANCE • differential diagnosis and severity of different types of anemia • Protein depletion and malnutrition • liver disorders. • chronic gastrointestinal bleeding • certain unusual disorders • iron poisoning • Thalassemia • hemosiderosis • hemochromatosis

26. PHYSIOLOGY • SERUM IRON (SI) • Men: 65 to 176 μg/dL • Women: 50 to 170 μg/dL • Newborns: 100 to 250 μg/dL • Children: 50 to 120 μg/dL • TOTAL IRON BINDING CAPACITY (TIBC) • Normal: 240–450 μg/dL • TRANSFERRIN SATURATION • Male 20–50% • Female 15–50% • FERRITIN LEVEL • Men: 18-270 (ng/mL) or 18-270 (mcg/L) • Women: 18-160 ng/mL or 18-160 mcg/L • Children: (6 months to 15 years) 7-140 ng/mL or 7-140 mcg/L • Infants: (1 to 5 months) 50-200 ng/mL or 50-200 mcg/L • Neonates: 25-200 ng/mL or 25-200 mcg/L • TRANSFERRIN • Normal: 204–360 mg/Dl

27. PATHOLOGY * = or normal

28. PRINCIPLE transferrin iron hydroxylamine Acetic acid thioglycolate Ferrous state Complexes immediately FerroZine Iron Reagent ferrous ion This change in absorbance is directly proportional to the concentration of iron in the sample. The system monitors the change in absorbance at 560 nm at a fixed-time interval

29. HAMEOGLOBINOPATHY • Oxygen transport metalloprotein • In mammals 97% of the RBC’s dry content • “Pathos” which means suffering • Suffex path is used to indicate disease like psychopath. Fatima Riaz

30. STRUCTURE • 2 Alpha and 2 Beta subunits- α2ß2 • Each associated with a heme group • Heme contains iron bound in porphyrin ring • Binds to Histidine 87 in α and Histidine 92 in β • Cleavage plane for Oxygen association. • Sequence closely related, difference of one AA in β and α chain of humans • and chimpanzees, variants in 7% of the world’s population. • Some produce harmless variants, while some produce hemoglobinopathies

31. Sickle Cell Disease • SCA or Drepanocytosis is an autosomal recessive genetic disorder which causes the cells to be rigid and crescent shaped. • Caused by a point mutation in the β-globin chain of hemoglobin on chromosome 11, causing the hydrophilic amino acid glutamic acid to be replaced with the hydrophobic amino acid valine. • The association of two wild-type α-globin subunits with two mutant β-globin subunits forms hemoglobin S (HbS) • Homozygous individuals are called HbSS and Heterozygous known HbAS

32. PATHOPHYSIOLOGY

33. DIAGNOSIS • Early diagnosis as part of screening tests is vital • Initial test by primary care doctor and successive other by hematologist

34. Presented by: Midhat Mustafa Thalassemia

35. Introduction • Thalassemia is a group of inherited disorders of hemoglobin synthesis characterized by defective globulin chains. • The name is derived from Greek words Thalasso= Sea and Hemia= Blood meaning anemia of sea. • Two types on basis of reduction of respective globulin chain • α- Thalassemia • β- Thalassemia • May involve other chains but alpha & beta are of prime importance. • Demography

36. Chromosome 16 Mild anemia Smaller blood cells that are lighter In color Very lightly colored red blood cells And possible severe anemia Death before birth (embryonic lethality)

37. Beta Thalassemia Inheritance • Mutations on chromosome 11 – 2 gene clusters • Beta thalassemia minor, also called thalassemia minor or thalassemia trait – Mild anemia • Beta thalassemia intermedia, also called thalassemia intermedia or mild cooley’s anemia – Bony deformities, fatigue,increased levels of iron in blood • Beta thalassemia major, also called thalassemia major or cooley’s anemia – jaundice, dark urine, protruding abdomen, defromed bones, fatigue.