Transcript red blood cell test

Routine Hematology
Tests
Group Members
Aqsa Abbasi
Ayesha Sajid
Azka Khan
Midhat Mustafa
Sadia Arshad
Fatima Riaz
Rida Rehman
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
Routine Blood
Hematolgy
Complete
Cell Tests
Count
Hemoglobin
value
Hematocrit
value
Red Blood Cell
Count
A complete blood count, often referred to as a CBC, is a common blood test.
Complete
A CBC provides detailed information
about three types of cells in the blood.
White Blood
Alexander
Vastem is
Cell Count
clinical purposes.
Platelet
the first Blood
person tocell
use the complete
blood Count
count for
count(CBC)
A CBC is routinely performed during annual physical examinations
Reticulocyte
count
Erythrocyte
sedimentataion
Rate (ESR)
Prothrombin
Time
Blood cell count principles
AUTOMATIC METHOD
• automated analyzer,
flowcytometry
• 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
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)
•Polycythemia vera
•Primary thrombocythemia
•Recent spleen removal
CBC (RBCs, WBCs)
Ayesha Sajid
RED BLOOD CELL TEST
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
HEMOGLOBIN
 Total amount of oxygen carrying protein inside RBCs
RANGE
Male 14-18 Gm/100 ml
Female 12-16.4 Gm/100 ml
HEMATOCRIT
 Percentage of blood volume made up of RBCs
RANGE
Male
42%-54%
Female 38%-46%
RED BLOOD CELL INDICES
Mean corpuscular
volume (MCV)
Mean corpuscular
hemoglobin
(MCH)
TYPES
OF ANEMIA
Mean corpuscular
hemoglobin
concentration (MCHC)
• Calculates the
size
• anemia:
Amount small
of
Microcytic
RBCs
of RBCs
per red blood
• Concentration of
Normocytichemoglobin
anemia: Normal
red blood cell
hemoglobin per
• Differential cell
red blood cell
diagnosis of Macrocytic
• Differential
anemia: larger than
anemias
diagnosis of
• Used for
normal red blood
cell
anemias
laboratory quality
control
TOO MUCH VS TOO LOW
HIGH
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 CELLS
Measures total white blood cell component 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
Leukocytosis
Leukopenia
• Acute infections
• Inflammation and
tissue necrosis
• Metabolic disorders
• Poisoning
• Leukemias
• Viral, bacterial,
parasitic infections
• Drugs
• Folate and vitamin
B12 deficiencies
• Bone marrow
infiltration
PRINCIPLE
Whole blood is
diluted 1 in 20 in an
acid reagent and
white blood cells are
counted
DIFFERENTIAL WBCs COUNT
LEUKEMIA
Types 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 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 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 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.
PRINCIPLE
• 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%
Prothrombin Time
Evaluating Blood Clotting Efficiency
Physiology- The Story Behind
Hemostasis – 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.
The time taken for the
blood to coagulate is
recorded. This is the
prothrombin time.
A chemical with anticlotting 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.
Reference Ranges
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
• Each laboratory should establish its own
normal range but in general the
prothrombin time for a normal plasma
sample, lies between 12-16 seconds
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.
RIDA REHMAN

five different types of tests
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Serum iron level test (SI)
total iron-binding capacity (TIBC) test
Ferritin tests
transferrin test
Transferrin saturation test
SIGNIFICANCE

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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

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 18270 (mcg/L)
 Women: 18-160 ng/mL or 18160 mcg/L
 Children: (6 months to 15
years) 7-140 ng/mL or 7-140
mcg/L
 Infants: (1 to 5 months) 50200 ng/mL or 50-200 mcg/L
 Neonates: 25-200 ng/mL or
25-200 mcg/L

TRANSFERRIN
 Normal: 204–360 mg/Dl
* = or normal
transferrin
iron
hydroxylamine
Acetic acid
thioglycolate
Ferrous state
Complexes immediately
ferrous ion
This change in absorbance is
directly proportional to the
concentration of iron in the
sample.
FerroZine Iron
Reagent
The system monitors the change
in absorbance at 560 nm at a
fixed-time interval
HAMEOGLOBINOPATHY
o
o
Oxygen transport
metalloprotein
In mammals 97% of
the RBC’s dry content
Fatima Riaz
o“Pathos” which means
suffering
o Suffex path is used to
indicate disease like
psychopath.
STRUCTURE
o 2 Alpha and 2 Beta subunits- α2ß2
o Each associated with a heme group
o Heme contains iron bound in
porphyrin ring
oBinds to Histidine 87 in α and
Histidine 92 in β
oCleavage plane for Oxygen
association.
Globins
Polypeptide
chains
Amino Acid
sequence
Genes
oSequence closely related, difference of one AA in β and α chain of humans
and 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 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
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
Presented by: Midhat Mustafa
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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
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)
Inheritance
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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.
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.