group members aqsa abbasi ayesha sajid azka khan midhat mustafa sadia arshad fatima riaz
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Group MembersAqsa AbbasiAyesha SajidAzka KhanMidhat MustafaSadia ArshadFatima RiazRida Rehman
Routine Hematology Tests
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 disorderstreating hematological malignaciesScreen, diagnose and measure the severity of diseasein the science of blood transfusion and the work of a blood bankin bone marrow and stem cell transplantation
Hemoglobin value
Hematocrit value
Red Blood Cell Count
White Blood Cell Count
Complete Blood cell
count(CBC)Platelet Count
Reticulocyte count
Erythrocyte sedimentataion Rate (ESR)
Prothrombin Time
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
Blood cell count principles
AUTOMATIC METHOD• automated analyzer, flow-
cytometry• It counts the numbers and
types of different cells within the blood.
• It aspirates a very small amount of the sample through the narrow tubing.
• light detectors and electrical impedance.
• One way the instrument can tell what type of blood cell is present is by size.
• Other instruments measure different characteristics of the cells to categorize them.
MANUAL BLOOD COUNT
• This measurement is made with a microscope and a specially ruled chamber(hemocytometer) using diluted blood.
• Counting chambers that hold a specified volume of diluted blood are used to calculate the number of red and white cells per litre of blood
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)•Polycythemia vera•Primary thrombocythemia•Recent spleen removal
•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
Platelet Count
CBC (RBCs, WBCs)
Ayesha Sajid
RED BLOOD CELL TESTNumber of RBCs per volume of whole blood
RANGEMale 4.5-6.4 Mil/c mmFemale 4.0-5.4 Mil/ c mm
HEMOGLOBIN Total amount of oxygen carrying protein inside RBCs
RANGEMale 14-18 Gm/100 mlFemale 12-16.4 Gm/100 ml
HEMATOCRIT Percentage of blood volume made up of RBCs
RANGEMale 42%-54%Female 38%-46%
RED BLOOD CELL INDICES
Mean corpuscular volume (MCV)• Calculates the
size of RBCs• Differential
diagnosis of anemias
Mean corpuscular hemoglobin (MCH)• Amount of
hemoglobin per red blood cell
• Differential diagnosis of anemias
Mean corpuscular hemoglobin concentration (MCHC) • Concentration
of hemoglobin per red blood cell
• Used for laboratory quality control
TYPES OF ANEMIAMicrocytic anemia: small RBCsNormocytic anemia: Normal red blood cellMacrocytic anemia: larger than normal red blood cell
TOO MUCH VS TOO LOWHIGH LEVELS LOW LEVELS
Dehydration Anemia
Polycythemia vera Autoimmune diseases
Kidney disease hemorrhage
heart or lung disease Bone marrow failure
Smoking Hemolysis
High altitude Long-term infections and poor diet and nutrition
WHITE BLOOD CELLSPRINCIPLE
Whole blood is diluted 1 in 20 in an acid reagent and white blood cells are counted
Measures total white blood cell component of whole bloodRANGEMales: 5,000 to 10,000 per microliter (µL) of bloodFemales: 4,500 to 11,000 per microliter (µL) of bloodLeukocytosi
s• Acute infections• Inflammation and
tissue necrosis• Metabolic
disorders• Poisoning• Leukemias
Leukopenia
• Viral, bacterial, parasitic infections
• Drugs• Folate and vitamin
B12 deficiencies• Bone marrow
infiltration
DIFFERENTIAL WBCs COUNT
LEUKEMIATypes of leukemia Description
Acute myelogenous leukemia (AML)
Cancer of myeloid cells. Too many WBCs and not enough RBCs and platelets.
Chronic myelogenous leukemia (CML)
increased and unregulated growth of predominantly myeloid cells.Increased granulocytes
Acute lymphocytic leukemia (ALL)
Characterized by excess lymphoblasts.
Chronic lymphocytic leukemia (CLL)
CLL affects B cell lymphocytes. Lymphocytosis
RETICULOCYTE COUNT
Azka Khan
Reticulocyte CountReticulocytes • 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 RNA that becomes visible under a microscope with certain stains such as new methylene blue.
•
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.
High valuesA 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 valuesA 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.
• Whole blood anticougulated with EDTA, is stained with supra-vital dyes (stain living cells) such as methylene blue.
• As reticulocytes are 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
Adults 0.5%-1.5%
Newborns Newborns:3%-6%
PRINCIPLE
Prothrombin TimeEvaluating Blood Clotting
Efficiency
Physiology- The Story BehindHemostasis – Prothrombin’s Role in Coagulation
Significance – Why the test is performed?
Evaluation of bleeding disorder- Easy bruising, excessive bleeding after dental appointment, blood in urine, heavy menstrual
cycle, etc.
Testing clotting ability before surgery
Monitor effects of warfarin treatment
Principle: How the test is performed?Blood is
removed from the patient.
A chemical with anti-clotting properties is added to prevent the
withdrawn blood from clotting (oxalated).
Calcium and tissue factor is then added to the blood in order to test the clotting time. This activates the
extrinsic pathway of coagulation.
The presence of calcium ions and tissue factor
should activate the relevant clotting factors to convert
prothrombin into thrombin.
The time taken for the blood to coagulate is recorded. This is the prothrombin time.
Reference Ranges
•Each laboratory should establish its own normal range but in general the prothrombin time for a normal plasma sample, lies between 12-16 seconds
Source of Tissue Factor e.g.
human, rabbit etc
- The exact technique used e.g. manual or
automated
Method of end-point
determination e.g. optical or
mechanical
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.
IRON PROFILERIDA REHMAN
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
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
PATHOLOGY * = or normal
PRINCIPLEtransferrin
Acetic acid
iron
Ferrous state
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
Complexes immediately
FerroZine Iron Reagent
ferrous ion
thioglycolate
hydroxylamine
HAMEOGLOBINOPATHY
o Oxygen transport metalloprotein
o In mammals 97% of the RBC’s dry content
o“Pathos” which means sufferingo Suffex path is used to indicate disease like psychopath.
Fatima Riaz
STRUCTUREo 2 Alpha and 2 Beta subunits- α2ß2 o Each associated with a heme groupo Heme contains iron bound in porphyrin ringoBinds to Histidine 87 in α and Histidine 92 in βoCleavage plane for Oxygen association.
GlobinsPolypep
tide chains
Amino Acid
sequence
Genes
oSequence closely related, difference of one AA in β and α chain of humansand chimpanzees, variants in 7% of the world’s population.oSome produce harmless variants, while some produce hemoglobinopathies
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 -globin chain βof 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 -globin subunits with two αmutant -globin subunits forms hemoglobin S (HbS)β
• Homozygous individuals are called HbSS and Heterozygous known HbAS
PATHOPHYSIOLOGY
DIAGNOSIS• Early diagnosis as part of screening tests is vital• Initial test by primary care doctor and successive
other by hematologist
Amniocentesis for prenatal diagnosis
Sickledex Tube Test
Hemoglobin Electrophoresis
ThalassemiaPresented by: Midhat Mustafa
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
Chromosome 16
Smaller blood cells that are lighter In color
Mild anemia
Very lightly colored red blood cells And possible severe anemia
Death before birth (embryonic lethality)
Beta Thalassemia
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.
Inheritance
Diagnosis:
• Complete blood count (CBC) and special hemoglobin studies.• Tests of the amount of iron in the blood• Family genetic studies, prenatal testing.
Treatment:
• Regular blood transfusions, iron chelation therapy, and bone marrow stem cells transplants
• Surgery of spleen or gall bladder.• Fetal hemoglobin therapy.
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