HLT41812 Certificate IV in

Pathology
HLTPAT410D Collect Pathology specimens other
than blood for specialized testing
HLTPAT411D Perform blood collection for
specialized testing

HLTPAT410D - Collect pathology specimens other

than blood for specialised testing

1.
2.
3.
4.

Prepare for specimen collection.

Perform procedure for specialised test
Perform post collection procedures.
Label and store specimens for
transportation.

HLTPAT411D Perform blood collection for

specialised testing

1. Prepare for blood collection

2. Perform collection for specialised test
3. Perform post-blood collection
procedures.
4. Label and store specimens for
transportation

Specialised tests
Circulatory and cardiac systems
As a part of these units you need to have
some understanding of basic anatomy
and physiology and any conditions that
require specialized tests to be performed
in the pathology collection rooms.
There are many disease processes which
you will come in contact with however
the following are some of interest.

Haemophilia
Haemophilia belongs to a group of
inheritable blood disorders that includes
haemophilia A, haemophilia B (Christmas
disease) and Von Willebrand's disease.
In haemophilia, the blood's ability to clot
is severely reduced because an essential
clotting factor is partly or completely
missing. This means that people bleed
for longer than normal.

Haemophilia A
Haemophilia A is a deficiency of an
essential clotting factor called factor VIII,
which is normally produced in the liver.
The faulty gene is found on the X
chromosome (X-linked). It's five times
more common than haemophilia B.

Hereditary factor
Every person has two sex chromosomes.
Women have two X-chromosomes while a
man has one X and one Y chromosome. Both
factor VIII and IX proteins are located on the X
chromosome.
A fault in the Factor VIII or IX gene on the X
chromosome will result in Haemophilia A or B
in males because there is not a normal X
chromosome to balance the abnormality.
Females carrying the haemophilia gene, the
normal X chromosome compensates for the
abnormal X chromosome.

Hereditary factor
An affected male cannot pass the disorder
on to his sons but all his daughters will carry
the haemophilia gene.
Females only rarely have symptoms of the
disorder, they may carry the haemophilia
gene and may pass on the disorder to their
sons.
Sons of women with the haemophilia gene
have a one in two chance of being affected
and daughters will have a one in two chance
of carrying the haemophilia gene.

Symptoms and long-term

problems
Symptoms range from easy bruising to
prolonged bleeding.
Bleeds can occur spontaneously (without
an external cause) or as a result of injury.

Symptoms and long-term

problems
How easily or badly a person bleeds
depends on the severity of their
deficiency. Minor cuts and grazes don't
usually cause any problems, but internal
bleeding can be life threatening, while
repeated bleeding in the joints typically
leads to arthritis or long-term joint
damage.

Symptoms and long-term

problems
Mobility problems may also result from
these spontaneous bleeds.
Haemorrhages into the brain are
particularly difficult to manage and can be
fatal.
Other long-term problems include the risk
of infection from blood products.
Education and employment
may also be disrupted

Diagnosis
Carriers of the gene can be identified with
a blood test.
Pregnant women can be assessed using
ultrasound to determine the sex of the
baby and so estimate the risk of the
disease.
They can then decide whether to have
more invasive tests - amniocentesis and
CVS - which can detect the relevant gene
in the baby.

Treatment
There's no cure for
haemophilia and,
although patients are
treated with injections of
the missing clotting
factor, there's no
permanent way of
replacing or increasing its
level.

Treatment
Haemophiliacs may receive medication,
such as desmopressin, to try to raise the
levels of the missing clotting factors,
especially prior to planned surgery or
dentistry, or they may receive factor
concentrate.
Management of acute bleeding remains an
essential part of their management if longterm health is to be sustained.
Severely affected children often receive
regular injections to prevent bleeding

Haemostatic Disorders
THROMBOCYTOPENIA
HAEMOPHILIA
Some tests you will deal with:
Fibrin degradation tests (FDP)
Prothrombin Time (PT)
Partial Thromboplastin Time/activated
partial Prothoplastin time (PTT/APTT)
Factor Assays

Tests
Fibrin degradation
tests (FDP)
Prothrombin Time
(PT)
Partial
Thromboplastin
Time/activated
partial
Prothoplastin time
(PTT/APTT)
Factor Assays

Look up your book:

What tubes would
you use?
How much Blood?
What department
do you send the
sample?
Are there any
special
requirements?

Blood Types
ANTIBODY
Any body or substance soluble or cellular
which is invoked by the stimulus provided
by the introduction of antigen and which
reacts specifically with the antigen in
some way

Blood Types
ANTIGEN
Any substance that as a result of coming
in contact with appropriate cells induces
a state of sensitivity and /or immune
response in some demonstrable way

Anemia
The site of the
bleeding must be
found and the
bleeding arrested.
Transfusion may be
necessary.

Tests for the presence of

Haemolysis
Laboratory findings
Positive or negative DAT (Direct Antiglobulin
Test) red cell abnormalities
Specimen Blood serum significance:
Corticosteroid therapy eg. Prednisolone
Haptoglobins: Haemolysis due to auto
immune disease, transfusion reaction,
mechanical trauma (heart valve) ineffective
red cell production due to severe Vit B12 or
folic acid deficiency. Overactive spleen, liver
disease.

Tests for causes of

Haemolysis
Further tests to identify the cause of
haemolysis:
Direct Antiglobulin Test /DAT
Direct Coombs Test / DCT
Used to separate haemolytic anaemia due to
an immune reaction from non-immune
causes a positive test detects the presence
of antibodies and therefore indicates an
immune cause.
Symptoms: Anaemia, jaundice, tea
coloured urine

Immunoglobulins
Anti bodies consists of chains of protein
attached to a sugar molecule
One part binds to antigens
One part interacts with elements of the
immune system such as neutrophils and
macrophages which have Fc receptors on
their surfaces
There are five classes of
immuniglobulins:IgG IgA IgM IgD

Tests for causes of

Haemolysis
Other laboratory results
Bilirubin Haptoglobins
Specimen Blood: Haematology
Method: tests for the presence of anti redcell IgG, IgA Antibodies or complement
attached to RBC, which cause RBCs to clump
together.

Cold aggluntins
A cold aggluntins is an antibody that
attaches to the red blood cell and causes
them to clump together or agglutinate at
a temperature below body temperature
Cold aggluntins are present in the serum
of person with mycoplasma pneumoniae
(an atypical pneumonia) glandular fever,
syphilis and certain blood diseases such
as haemolytic anaemia

Cold aggluntins
To prevent the antibody from attaching
to the RBCs, a cold agglutinin specimen
must be collected in a tube pre-warmed
to 37 and kept at this temperature until
the serum is separated form the cells
(look up special instructions).

Coagulation Studies
Excessive bleeding may result from:
Inability to arrest blood loss ( Platelet
count or platelets that do not function
properly).
Failure to form a permanent clot
(defect in the clotting process).
Overactive of the system that breaks
down clots (excessive fibrinolysis).
Easy breakage in the smallest blood
vessels (fragile capillaries).

Coagulation Studies
There are two alternative pathways:
Intrinsic - stimulated by contact with a
foreign surface or material
Extrinsic - activated by tissue damage.
The final two reactions is the conversion
of prothrombin to thrombin and fibrinogen
to fibrin.

Tuberculosis
An illness caused by a
germ mycobacterium
tuberculosis The lung is the organ
commonly infected.
The germ may occasionally infect the
kidneys, bones and joints, lymph
glands and other parts of the body.

Tuberculosis
About 800 cases are found and treated
in Australia each year
Newly arrived immigrant who caught the
disease prior to arrival are at risk
More likely to affect people with low
resistance to infection, mal-nourished etc.

Tuberculosis
Only 5 10 % of people who become
infected with TB get sick.
TB can lie dormant for many years and
when the immune system is weakened the
chance of getting sick increases.
Dissemination of tuberculosis is when the
infection spreads from the lungs to other
organs.

Tuberculosis
SYMPTOMS
Fever
Cough
Loss of energy
tiredness
Nocturnal sweats
Weight loss
Phlegm may
sometimes have
the presence of
blood

TREATMENT
3-4 antibiotics
taken together over
many months to be
certain of a cure
Time will vary
according to many
factors
Follow up regularly

Insert Digestive
in here

AFB Smear
AFB (shown in red) are tubercle bacilli

AFB (shown in red) are tubercle bacilli

Also known as: TB culture and sensitivity
Formal name: Acid-fast bacillus smear and culture and sensitivity

Tuberculosis

Tuberculosis

Tubercular Lesion

Klebsiella Pneumoniae
The patient initially
improved with antituberculosis
treatment but then
developed new
fever and cough.
Sputum cultures
were positive for
Klebsiella
pneumoniae.

A 50 year old female presented

with four weeks of cough

Diagnosis
The most useful
diagnosis methods
are:
Chest x-ray
Sputum cultures
Tuberculin skin
test
Bronchoscopy
Open lung biopsy

QuantiFERON TB Gold
Blood Collection
There are three tubes:
1. Red Cap = TB Antigen
2. Grey Cap = Nil negative control
3. Purple cap = Mitogen control

Collection of blood
1. Collect 1ml exactly into each tube
Blood tube MUST be filled to 1ml for
optional test performance ie to
INDICATOR line
Tubes may fill slowly, keep tube on
needle until blood flow stops to
ensure correct volume
For butterfly needle always use a
PURGE tube

Collection of blood
2. Shake tubes for 5 seconds to resuspend
ingredients on the tube wall
3. Do NOT refrigerate. Leave at room
temperature (maximum 16 hours)

BCG Vaccination

Tuberculin PPD
Contains soluble growth products derived
from the tubercle bacillus. When
administered intra-dermally either by
injection or by means of a puncture
devise, a hypersensitivity reaction,
manifesting as induration as erythema
will appear in sensitive individuals.

Mantoux testing procedure

using Tuberculin PPD
Inject intradermally 0.1ml of a solution
containing 1.00IU per ml (ie 10IU per
dose of 0.1ml) into the ventral surface of
the upper part of the forearm.
The reaction commences within 24 hours
and reach maximum size in 48- to 72
hours.
The result should be read in 72 hours but
could be read from 48 hours to the fifth
day.

Mantoux testing procedure

using Tuberculin PPD

Mantoux testing procedure

using Tuberculin PPD
A positive reaction is an indication that
the patient has/had at some time a
tuberculosis infection. A positive test
does not indicate an active infection, but
indicates that further investigation should
be done.

Mantoux testing procedure

using Tuberculin PPD
Tuberculin skin test
involves injection of
PPD ( a purified
protein derivative)
into an area like the
forearm (pictured
right). If there is a
reaction, TB is
present

Mantoux testing procedure

using Tuberculin PPD
It is the oedema and induration that is
important and can usually be detected
with a finger rather than the eye.
he diameter of the area of induration or
oedema is measures in millimeters and
recorded.
Erythema without oedema or induration
should be disregarded.

Mantoux testing procedure

using Tuberculin PPD

Reading the Tuberculin

skin test
Read reaction 48-72
hours after injection
Measure only
induration
Record reaction in
millimeters

Mantoux testing procedure

using Tuberculin PPD
The National Tuberculosis Advisory Council
(Canberra) has suggested the following
degrees of reaction to the 10U dose
Mantoux Test:
Negative: less than 5mm diameter
Weak positive : 5-9mm diameter
Intermediate positive: 10-14 diameter.
Strong positive : 15mm diameter or more
vesiculation

Prevention and treatment

Routine BCG vaccination occurred in
Victoria from the early 1950s until 1984
In the US routine skin testing is done
during baby check-ups
Anyone who has been exposed to TB
should have a tuberculin skin test
Infected people should stay home from
work or school to prevent spreading TB

Prevention and treatment

TB can be treated using drugs like:
o Isoniazid (INH)
o Rifampin
o Pyrazinamide
o Ethambutol
o Streptomycin
People with TB take a combination of drugs.
MDR (Multi-drug-resistant) TB develops
when people do not follow instructions about
medication.

Tuberculosis Facts
TB is starting to become more common
again
HIV is speeding up the spread of TB
TB accounts for about 15% of deaths of
HIV sufferers worldwide
Greater movement of people around the
world is helping to spread TB
In the US, nearly 40% of TB cases are
among foreign born people

Tuberculosis Facts
Untreated TB can spread quickly through
refugee camps
Up to 50% of the worlds refugees may be
infected with tuberculosis
In 1995 about 30% of San Franciscos
homeless were infected with TB
compared to an overall 7% of the US
population

Tuberculosis Facts
Kills more people in the world than any
other infectious disease
Has probably been around for more than
2000 years
Has been known by names such as:
o Phthisis
o Consumption
o White plague

Tuberculosis Facts
In Victoria the
Western
Metropolitan area
has the largest
incidence
This is primarily
due to the number
of immigrants from
Asia, Africa,
Europe and the
Middle East

Of immigrants,
males 25 29 are
the highest risk
group
Changes to Visa
procedures for
overseas students
now require them to
undergo a medical
examination before
approval

Tuberculosis Facts
Only people with
pulmonary (lung) TB
are infectious
An untreated person
with infective TB will
infect 10 15 people
each year
A person only needs
to breathe in a few
bacteria to become
infected

Tuberculosis Diagnosis
A chest x-ray shows lungs, heart and
diaphragm. A chest x-ray can be ordered
for any chest complaint,
Sputum cultures identify bacteria in a
sample of sputum from the lungs
A Bronchoscopy is when a camera is
sent down the throat into the lungs so the
doctor can see what is happening inside
the lungs

Underactive adrenal glands/

Addisons Disease
Addison's disease is the result of an under
active adrenal gland. An under active adrenal
gland produces insufficient amounts of
corticosteroid hormones. Four in every
100,000 people have Addison's disease.

Causes
Most of the time, the cause of the disease is
unknown. About one-third of Addison's
disease cases are caused by the actual
destruction of the adrenal glands through
cancer, infection, or other diseases. Other
causes may include:
Use of corticosteroids as a treatment (such
as prednisone) causes a slow down in
production of natural corticosteroids by the
adrenal glands.
Certain drugs used to treat fungal infections
may block production of corticosteroids in
the adrenal glands.

Signs and symptoms

weakness
fatigue
dizziness
dark skin
black
freckles
weight loss

dehydration
lack of appetite
muscle aches
nausea
vomiting
diarrhea
intolerance to
cold

Signs and symptoms

bluish-black
discoloration around
the nipples, mouth,
rectum, scrotum,
or vagina

Diagnosis
In addition to a complete medical history
and medical examination, diagnostic
procedures for Addison's disease may
include:
blood tests to measure corticosteroid
hormone levels
kidney function tests to determine if urine
is concentrated

Treatment
Since Addison's disease can be life threatening,
treatment often begins with administration of
corticosteroids. Corticosteroids, such as
prednisone, may be taken orally or
intravenously, depending on the patient's
condition.
Usually the patient has to continue taking the
corticosteroid the rest of his/her life. Treatment
may also include taking fludrocortisone, a drug
that helps restore the body's level of sodium and
potassium.

Overactive adrenal glands/

Cushings Syndrome
Cushing's syndrome is the result of the
excessive production of corticosteroids by the
adrenal glands. An overproduction of
corticotropin -- the hormone that controls the
adrenal gland -- by the pituitary gland, which
stimulates the adrenal glands to produce
corticosteroids, may be one cause. In addition,
certain lung cancers and other tumors outside
the pituitary gland may produce corticotropins.

Cushings Syndrome
upper body obesity
round face
increased fat around
neck
thinning arms and
legs
fragile and thin skin

Cushings Syndrome
stretch marks on
abdomen, thighs,
buttocks, arms, and
breasts
bone and muscle
weakness
severe fatigue
high blood pressure
high blood sugar

Cushings Syndrome
irritability and
anxiety
excess hair growth
in women
irregular or stopped
menstrual cycles in
women
reduced sex drive
and fertility in men

Diagnosis
In addition to a complete medical history and
medical examination, diagnostic procedures for
Cushing's syndrome may include:
X-rays to locate any tumors
24-hour urinary test to measure for corticosteroid
hormones
Computed tomography (CT or CAT scan) - a noninvasive procedure that takes cross-sectional
images of the brain or other internal organs; to
detect any abnormalities that may not show up on
an ordinary x-ray
Magnetic resonance imaging (MRI) - a noninvasive procedure that produces two-dimensional
views of an internal organ or structure, especially
the brain or spinal cord

Diagnosis
Dexamethasone suppression test - to
differentiate whether the excess production
of corticotropins are from the pituitary gland
or tumors elsewhere
corticotropin-releasing hormone (CRH)
stimulation test - to differentiate whether the
cause is a pituitary tumor or an adrenal
tumor
Other laboratory tests

Synacthen Stimulation Test

Synthetic ACTH, known as synacthen, is used to
stimulate the adrenal cortex to produce cortisol
METHOD: No preparation is necessary.
Blood sample taken to measure baseline
cortisiol.
An injection of synacthen is administered
Blood samples are taken at 30 minutes and 60
minutes
SIGNIFICANCE: Failure to raise cortisol above
test baseline indicates adrenal insufficiency

Dexamethasone
Dexamethasone, a corticosteroid, is similar to
a natural hormone produced by your adrenal
glands. It often is used to replace this
chemical when your body does not make
enough of it.
Relieves inflammation (swelling, heat,
redness, and pain)
Used to treat certain forms of arthritis; skin,
blood, kidney, eye, thyroid, and intestinal
disorders (e.g., colitis); severe allergies; and
asthma.
Dexamethasone is also used to treat certain
types of cancer.

Dexamethasone
Suppression test
In Cushings disease and ACTH producing
cancers (eg certain lung cancers or other
tumours) the raised cortisol level is not
suppressed by Dexamethasone.
Measurement of cortisol forms the basis of
both this and the Synacthen Stimulation test
Specimen: Blood

Dexamethasone
Suppression test
METHOD:
Dexamethasone is taken by tablet at
midnight : Cortisol measures from a blood
sample collected 8 hours later (8am)
NORMAL VALUES:
Check with laboratory, normally 200-650
nmol/L cortisol

Dexamethasone
Suppression test
SIGNIFICANCE:
Non suppression of cortisol with a dose of
Dexamethasone - Cushings Disease
High dose Dexamethasone non suppression
ACTH-producing lung or stomach cancer,
adrenal tumour or cancer (high doses of
Dexamethasone will usually suppress the
pituitary tumour production of ACTH in
Cushings disease)
FURTHER TESTS:
CT, MRL, Insulin Hypoglycaemic
Stimulation Test, Synacthen Stimulation Test.

Treatment
Treatment for Cushing's syndrome
depends on its cause.
Surgery may be needed to remove
tumors or the adrenal glands.
Other treatment may include radiation,
chemotherapy, and use of certain
hormone-inhibiting drugs.

Diabetes
There are three main types of diabetes that
require clinical care by a physician or other
healthcare professional:
1. type 1 diabetes
2. type 2 diabetes
3. gestational diabetes

Type I Diabetes
Type 1 diabetes is also known as diabetes
mellitus, insulin-dependent diabetes mellitus
(IDDM), juvenile diabetes, brittle diabetes, or
sugar diabetes. There are two forms of type 1
diabetes:
idiopathic type 1 - refers to rare forms of the
disease with no known cause.
immune-mediated diabetes - an autoimmune
disorder in which the body's immune system
destroys, or attempts to destroy, the cells in the
pancreas that produce insulin. Immunemediated diabetes is the most common form of
type 1 diabetes

Causes
The cause of type
1 diabetes is
unknown, but it is
believed that
people inherit a
tendency to
develop diabetes,
and that viruses
may be involved.

Causes
This auto-immune disease results from
the body's failure to produce insulin, the
hormone that allows glucose to enter the
cells of the body to provide fuel.
This is the result of an autoimmune
process in which the body's immune
system attacks and destroys the insulin
producing cells of the pancreas.

Causes
When glucose cannot enter the cells, it
builds up in the blood and the body's cells
literally starve to death.
People with type 1 diabetes must take
daily insulin injections and regularly
monitor their blood sugar levels

Signs and Symptoms

high levels of sugar in the blood when
tested
high levels of sugar in the urine when
tested
unusual thirst
frequent urination
extreme hunger but loss of weight
blurred vision
nausea and vomiting
extreme weakness and tiredness
irritability and mood changes

Diagnosis
Fasting plasma glucose of greater than or
equal to 126 mg/dl with symptoms of
diabetes.
Casual plasma glucose (taken at any time
of the day) of greater than or equal to 200
mg/dl with the symptoms of diabetes.
Oral glucose tolerance test (GTT) value of
greater than or equal to 200 mg/dl
measured at a two-hour interval. The GTT
is given over a two -hour time span.

Treatment
People with type 1 diabetes must have daily
injections of insulin to keep the blood sugar
level within normal ranges. Other parts of the
treatment protocol may include:
appropriate foods to manage blood sugar
level.
exercise to lower and help the body use
blood sugar.
regular blood testing for blood-sugar levels.
regular urine testing for ketone levels.

Type 2 Diabetes
Type 2 Diabetes
This type is far more common than type 1,
affecting between 90 percent and 95
percent of people with diabetes over age
20. It occurs when your body is resistant to
the effects of insulin or your pancreas
produces some, but not enough, insulin to
maintain a normal glucose level.

Blood glucose
Blood sugar levels (BSL)
Test used to measure glucose sugar in
the blood. It is oxidised be cells to
liberate energy for metabolic function.
The brain an nervous system rely almost
entirely on glucose for energy.
Specimen Oxolate. Specimen may be
random (taken approx. 2 hours after a
meal_ or fasting (taken at least 8 hours
after a meal)

Significance
Results should be interpreted in
conjunction with blood glucose results
Blood Glucose
Blood Sugar
Common Diabetes Common - low blood
mellitus
sugar (hypoglycaemia)
due to fasting missed
Uncommon - Excess
meals carbohydrate/
thyroid Hormone,
acute or chronic
insulin reactions
pancreatitis, mumps, Uncommon tumours of
the islets cells. Glucagon
cystic fibrosis, cancer
defeciency, cancer of
of the pancreas,
the adrenal glands, liver
stroke, heart attack,
chronic liver disease
damage due to
poisoning decrease
pituitary or thyroid
hormones

Glucose Tolerance Test

(GTT)
Used to confirm Diabetes Mellitus
Booked test - required to stay in the
rooms for 2 hours
Fasting test 8 hours
Dietary Requirements 3 days prior to
test patients diet should include approx.
15G of carbohydrate
Test Oxalate tubes . A standard dose of
glucose is given in a drink and blood
glucose levels are checked over the next 2
hours

Glucose Challenge Test

(GCT)
Used to screen pregnant women for
gestational Diabetes:
Specimen- Oxalate tube
Dietary requirements nil
Fasting no
Test - is performed at 26-28 weeks
gestation
Women are given a glucose drink and blood
specimen is collected 1 hour later

Chain of custody
TESTING:
Urinary Drug
screen
Alcohol levels
Paternity testing
Arsenic testing

REQUIREMENTS:
Identify in the
usual manner with
referral
Pre-label sample
container

Urinary drug screen

Accompany client to the toilet
Explain the procedure with emphasis that
it is urinary drug screen
Eg. I am required to see you pass urine
under the conditions of the screen
Two personelle at the branch
Do not put yourself in a position of being
compromised

Urinary drug screen

Chain of custody form filled out partly
before starting then signed on completion
of sample being obtained
If the client is unable to sign a designated
person to sign Guard or police officer to
verify signature .
Sample collection taken
Just an ordinary sample. A urinary drug
screen is not a MSU

Urinary drug screen

After collection:
In the presence of the client, seal the jar
with red tape or appropriate sealer with
the signature over the seal
Form is signed by the client or
designated signatory and verified
Specimen sent in a tamper proof
biohazard bag
If the laboratory does not receive it intact,
the specimen needs to be recollected

Urine Catheter collection

C.S.U= catheter specimen of urine.
Samples of urine collected from the
bladder catheter. Collected by attending
medical staff. Transported and stored a
4 centigrade