Chapter-3

Collection, handling and shipment of

laboratory specimens

By Yitayal.A (Bsc,
Msc))

1
Objective

After completion of session, the student will be

able to:
Define stool, blood and urine specimens

Identify stool, blood and urine specimens

Familiarize how to collect stool, blood and urine

specimen

Know specimen rejection criteria

Label and transport different types of specimen

2
Outline

3.1. Definition

3.2. Basic Concept in Specimen Collection

3.3. General Rejection Criteria

3.4.Guideline in specimen collection

3.4.1.Stool collection

3.4.2. Blood collection

3.4.3. Collection of sputum specimen

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3.1: Definition

Definition of specimen
o Specimen is a part taken to determine the
character of the whole.
Safety during collection, processing &
transportation

o Masks (e.g. sputum)

o Gloves

o Protective eye ware (goggles)

o Protective clothing (gown) 4

Specimen collection..
Several different kinds of specimens are used
routinely in the clinical laboratory.
These are:
Stool

Blood

Sputum

Urine

Pus (discharge)

Throat, eye, ear, vaginal swabs and others

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Specimen collection

o Correct treatment depend on accurate result

o Accurate result depend on quality specimen

o Quality specimen depend on proper :

Collection

Transportation

Processing of specimens

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Specimen collection

o Purpose of quality specimen:

provide high yield positive result

Provide that a negative result indicates the

absence of the pathogen.

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Specimen collection..
Importance of Quality Specimen on Patient Care
o Key to accurate laboratory diagnosis.
o Directly affects patient care & patient outcome.
o Influences therapeutic decisions.
o Impacts hospital infection control.
o Impacts patient length of stay, hospital costs,
and laboratory costs.
o Influences laboratory efficiency.
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3.2 Basic Concept in Specimen Collection

1. Site selection
a. Clinician
Should select appropriate tests & specimens based
on:
- Physical examination (sign & symptoms)
- Radiological examination
b. Laboratory personnel
Should collect specimens from actual infection site
with little external contamination by using :
Aseptic technique
To prevent contamination of specimen &
To protect the patient from infection
Sterile container
. Should collect specimens from right site

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Basic Concept in Specimen Collection

Approaches to Avoid contamination :

o Careful patient education

There are occasions when patients participate

actively in specimen collection (e.g. sputum,
urine).

Therefore, they must be given full instructions

& cooperation by the care giver.
o Educating the clinicians
How to collect & transport specimens through10
Specimen collection..

2. Volume of specimens

Collecting & processing too little specimen will

give us lower sensitivity.

Collecting adequate volume:

o enhance recovery of the pathogen.

o enable to perform all procedures required

or to permit complete examination.
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Specimen collection..

For example;

o Sputum: 5 -10 ml for mycobacterium examination.

o Blood:

Serology: minimum 2 - 3 ml.

Culture: 10 20 ml (adult) & 1-5ml

(infant).

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Specimen collection..

3. Time of collection

Provide best chance of recovery of the causative

agent .

Sputum & urine

early in the morning soon after the patient

awaken.

Blood

when the patients temperature begins to rise.13

Specimen collection..

4. Collect specimens before the administration of

antimicrobial

Because antimicrobials limits recovery of

pathogens.

5. Age of specimens

o Age of the specimen directly influences the

recovery of protozoan organism.
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Specimen collection..

6. Labeling
Make sure that you are collecting/drawing the right
person first.
Then label with:
o Patient name
o Unique identification number
o Time of collection
o Specimen collection date
o Specimen collection location
o Patient demographic information
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Specimen collection..

During Labeling:
o Make sure that container label & the requisition
match.
o Ensure the labels on the containers are adherent under
refrigerated conditions.

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3.3 General specimens Rejection Criteria
1. Unlabeled Specimens

o Common specimen like blood, urine, swabs, sputum,

stool, can be easily recollected.

o less common specimens like CSF, fluids, tissues,

etc. are more difficult to recollect.

Call the person who collected it for the

identification of the specimen.

o If he is unable to identify the specimen, the

ordering physician will be notified. 17
General Rejection Criteria

2. Incorrectly labeled (mislabeled) specimens

o Use same criteria as for Unlabeled Specimens.

3. Incorrect container or Preservative

o Specimens received in an incorrect container, or

without appropriate preservative, will require
recollection.

o So the patient will be contacted to arrange for

recollection of the specimen. 18
General Rejection Criteria

4. Insufficient specimen for procedure

o If insufficient, recollect (urine, stool, sputum,

blood, etc.)

o If the specimen is not recollectable (CSF, fluids,

etc.), the physician will be contacted to establish
a priority order of tests.

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General Rejection Criteria

5. Unsuitable Specimen for Procedures

Specimens which are unsuitable for the procedure
requested saliva for sputum test or specimen too
long for a valid result.

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Specimen Transportation
Required when:
Specimens are to be sent to referral laboratory.

o For teaching purpose.

o For Quality assurance.

o Unavailability of trained personnel around the

collection site.
o Specimens are collected in the field.
o Lack of time to examine within the recommended time
due to laboratory workload. 21
Specimen Transportation......

This transportation is made by using different

preservation methods

Physical

chemical.

- chemical method of preservation is most

common.

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Specimen Transportation......

Purpose of preservation :

o Maintain protozoan morphology

o Prevent development of helminthes eggs & larvae

o Maintain viability of microorganisms

o Prevent overgrowth of normal flora

o Prevent instability of solutes & degeneration of

sediments (e.g. urine)
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 Specimen Transportation......

Specimen packaging

o Screw container tops on firmly

o Wrap in absorbent wadding to absorb any fluid

leakage

o Place in a self-sealing plastic bag

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Specimen Transportation......

o Place the request form into the secondary pocket

of the specimen bag.

o Hazard labels with internationally accepted

biohazard label (HIGH RISK).

o Pack specimens in strong cardboard box or a

grooved polystyrene box.

o Seal with self-adhesive tape.

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Specimen Transportation......

Specimen Storage (Prior to Dispatch)

o Blood samples should be kept at +4 oc.

o Serum & plasma can be stored either frozen or at

+4oc.

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 Specimen Transportation......

Note: If they are frozen, they should not be unfrozen

until dispatch to the laboratory.

Transport

o Transport with labelled, tightly fitted, leak

proof container.

o Wet ice or ice pack should be used.

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3.4. Guideline for specimen collection

3.4. 1. Stool collection

Instruct the patient
To avoid contamination from:
Urine - may kill the pathogen.

Water - may contain free-living organism .

Proper sampling
Material from inside & surface of the feces.

Material which contain mucus & blood.

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Guide line

Adequate quantity

Tea spoon full (5ml) to large tea spoon full

(10ml).

Use: prevent rapid drying & permit detection of

parasites in low concentration.

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 .3.4.2.Blood collection
- Blood represents a large percentage of the total specimens
used in laboratory determinations.

Collection of Blood Specimens

There are three general sources of blood for clinical laboratory
tests:-
- Venous blood
- Peripheral, or capillary blood
- Arterial blood ( rare cases like in blood gas analysis)

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Blood collection.
1. Collection sites:
a. Capillary blood
o Ring/middle finger - adults & children.
o Heel - infant < 3 months side of heel.
o Big toe older infants (> 3 months) side of great
toe.
o Ear lobe today is not routinely used as a blood
collection site.

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Capillary blood

The capillary blood is obtained by skin puncture. It

provides only small quantities of blood specimen
for making a blood smear (differential count), cell
count or hematocrit determination.

Skin puncture specimen is preferred over

venipuncture specimen for the study of blood smear
and manual differential count.

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Blood collection by finger puncture
procedure

1. Assemble the necessary equipment, lancet alcohol

pad, dry surgical gauze, capillary tube,
microscope, slide and other supplies(glass, ,
marking penile, lead panicle, etc).

2. Be sure that the patient is seated comfortably.

3. Find a spot on the middle or ring finger of the left

hand. The spot is located on the side of the figure,
which is less sensitive than the tip.
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Finger puncture procedure

4. Clean the site with a sterile cotton wool swab dipped

in 70% alcohol, then remove the alcohol with a dry
sterile cotton wool swab. This remove dirt, and
epithlial debris. Warm up the part chosen site for
pricking, increase the blood circulation. And leave
the area relatively sterile.

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Finger puncture procedure

5. Grasp the figure firmly and make a quick , firm

puncture with a sterile lancet (sharp pointed blade).
The puncture should be 2-3 millimeter deep at the pre
located spot on the side of the figure in line with the
figure print striations.

- If a good puncture has been made , the blood will

flow freely. If it does not , use gentle pressure to
make the blood form a round drop. Excessive
squeezing will cause dilution of blood with tissue
fluid.
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Finger puncture procedure
6. Wipe away the first drop of blood with sterile cotton
wool. The first drop of blood may be contaminated
with tissue fluid and will interfere with the
laboratory result if used. The succeeding drops are
used for test.

7. Collect the specimen by holding a capillary tube to

the blood drop (for hematocrit determination), or by
sucking in to the Sahli pipette for the hemoglobin
determination and for blood count, or by touching the
drop to the glass slide for preparing smear.

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Blood collection.

b. Collection of venous blood

The volume of blood obtained by venipuncture is

sufficient to carry out multiple test.

Venipuncture can be done either by the syringe method

or vacuum tube method.

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Venous blood is

Venous blood is collected from a vein.

Veins in the forearm are most commonly used for

vein puncture.

Veins in the wrist or ankle may also be used for

vein puncture (if the forearm site is not
available).

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Venous blood

The three main veins in the forearm are:

Cephalic

Median

Basilic.

- The median cubital vein is usually chosen for vein

puncture. (Because it is larger, closer to the
surface, easier to enter).

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Veins of fore arm

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Blood collection.

Equipment and Supplies

Gloves
Needle
Holder
Tubes
Gauze
Alcohol Pad
Band Aid
Sharps
Container
Tourniquet

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Blood collection.

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Needle Gauge
Gauge is the diameter of the needle
The smaller the gauge, the larger the
diameter.
Routine adult collections use the 21
or 22G.
23G is used for children and
smaller, fragile veins.
25G may be used for scalp vein
draws on neonates and premature
infants.
Smaller gauge needles used with
full draw evacuated tubes may
cause pain, slow blood delivery., 43
and hemolytic.
Blood collection procedure by
venipuncture

1. Assemble all the things required during blood

collection

2. Read carefully the patients form, identify the

patient and decide patient and decide the total amount
of blood needed for the entire test.

3. Select the blood collection container and label them

with the patients identification number.

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Venipuncture procedure

4. Introduce your self to the patient. Ask the patient to

sit alongside the table used for taking blood. Lay
his arm on the table, palm upwards.

- The procedure of blood collection should be

explained by the vein puncturist to the patient to
minimize apprehension.

- Never draw blood from standing patient.

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Venipuncture procedure
5. Select the puncture site carefully after inspecting
both arms.

6. Apply the tourniquet before drawing blood. The

tourniquet should not be left in place unless the
technician is ready to proceed immediately with the
veinpuncture.

7. Using the index figure of your left hand , feel for the
vein where you will introduce the needle.
8. Disinfect the site with a swab dipped in methanol or
70% alcohol. Rub the venipuncture site thoroughly.
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Venipuncture procedure

9. Remove the syringe from the protective warp or test

tube used during sterilization and the needle from the
sterilized vial, assemble them and see the needle is
fixed tightly.

10. Puncture the vein, try to enter the skin first and
then the vein , at a 30 to 40 0 angle. Continue with
draw the position and fill the syringe with the
request amount of blood.
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 Venipuncture procedure

11. Release the tourniquet by pooling on the looped end.

12. Place a swab of cotton wool over the hidden point of the
needle. With draw the needle in one rapid movement from
under the swab.

13. Ask the patient to firmly on the cotton wool swab for 3
to 5 minutes. This stops bleeding from the wound. Do not
bend the arm , this may cause hematoma.

14. Remove the needle from the syringe and gently expel the
blood in to appropriate container. 48
 Venipuncture procedure
15. Mix the blood immediately and thoroughly but gently
with the anti coagulant. Label the bottle clearly with
the name of the patient, date, sex and registration number.
16. Immediately discard the syringe and the needle in
appropriate waste disposal equipment.
17. Before the patient leaves , re inspect the venipuncture
site to ascertain that the bleeding has stopped. If the
bleeding has stopped , apply an adhesive tap over the
cotton wool swab on the wound , otherwise continue to
apply pressure until the bleeding stops. Do not leave
the patient until the bleeding stops.

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Sites to Avoid for Venipuncture

Scarred areas

Hematoma

An arm with an IV

If unavoidable draw distal to IV

Edematous areas

Arm in which blood is being transfused

Arms with fistulas or vascular grafts

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3.4 .4.Collection of sputum specimen
Definition
Saliva: - is secreted by the salivary glands and
is limited in the oral region.

Sputum: - is the material coughed up from the

throat and lungs.

Sputum is usually examined to determine the

presence of disease of the lungs or of the upper
respiratory tract e.g. diagnose pulmonary
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Sputum.
Collection Method
Provide the patient with a sterile wide- mouthed
glass bottle of about 50 ml capacities with a
screw top.
he/she should take a very deep breath, filling
his/her lungs.
She/he should empty his/her lungs in one breath,
coughing as hand and deeply as she/he can.
She/he should spit what he brings up into the jar.

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Sputum..

After Collection
Check that a sufficient amount of sputum has been
produced.
The sputum of an infected person usually contains:
Thick mucus with air bubbles

Threads of fibrin

Patches of pus

Occasional brownish streaks of blood

N.B. The first sputum coughed out in the early

morning is the most desirable specimen for the
laboratory investigation. Why? 53
Sputum..

Liquid frothy saliva and secretions from the nose

and pharynx are not acceptable. Have the patient
produce another specimen.
Dispatch of sputum: - Fluid transport medium
A wide-mouthed, screw-top bottle containing: 25ml
of a solution of 0.6% cetylpyridinium bromide in
distilled water.
The patient should expectorate directly into the
liquid in the bottle.
Screw on the top and dispatch.

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Sputum..

55
QUALITY ASSURANCE
To deliver quality result lab personnel perform
the internal and external quality assessment
teqiniques.

Internal quality assessment for sputum,blood

and stool and other during specimens
collections are include like
checking the volume of specimen,
Checking the time of collection,

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Cont.
proper labeling of the container
Proper storage and transpot of specimens and
others
External quality controls methods are
Onsite supervision
Panal test
Blind rechecking

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Summary question

1. Explain how to collect, handle and transport stool, urine

and blood laboratory specimens.
2. what are the criteria to assess the quality of
specimens?

58
References
1. Linne Jean Jergenson, Basic techniques of medical
laboratory 4th ed. 2000.
2. WHO, Manual of basic techniques for a health
laboratory 2000.
3. Chees brough M.District Laboratory manual for
tropical courtiers, Cambridge Univerity press, 2000
(Vol. I ).
4. Chees brough M.District Laboratory manual for
tropical courtiers, Cambridge Univerity press, 2000
(Vol. II).
5. Seyoum B. Introduction to medical laboratory
technology students lecture note series 2002.
6. www.CDC.gov 59
THANK YOU

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