Chapter-3

Collection, handling and shipment of laboratory

specimens

By: Tegenaw Tiruneh (BSc, MSc,

Assistant professor)

1
Objective

After completion of this chapter, the student will be

able to:
 Define different types of specimen

 Identify different types of specimen

 Familiarize how to collect different types of 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

[Link] in specimen collection

[Link] collection

3.4. 2. Collection of Urine

3.4.3. Blood collection

3
Out line

3.4.4. Collection of sputum specimen

3 .4.5. Throat, eye, ear, Vaginal and urethral swab

collection

3.4. 6. Collection of Cerebro-spinal fluid (CSF)

3.4.7. Skin specimen

3.5. [Link] and transport of semen

4
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) 5

Specimen collection…..

◼ Several different kinds of specimens are used routinely

in the clinical laboratory.
◼ These are:
◼ Blood

◼ Urine

◼ Stool

◼ Sputum

◼ Pus (discharge)
6
Specimen collection…..

◼Throat, eye, ear, vaginal swabs

◼ Skin snips

◼ Body fluids (pleural, pericardial, peritoneal, synovial,

and cerebro spinal)

◼Hair, nail, coetaneous tissue

◼Biopsy tissue from an organ.

7
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

8
Specimen collection……

o Purpose of quality specimen:

✓ provide high yield positive result

✓ Provide that a negative result indicates the

absence of the pathogen.

9
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.
10
3.2 Basic Concept in Specimen Collection
1. Site selection
a. Clinician
• Should locate right anatomic site & 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
11
Basic Concept in Specimen Collection …..

▪ Sites of Infection where the Specimen is Likely to

Become Contaminated During Collection

o Lower respiratory tract ----- Oro-pharynx

o Bladder ……………..… Urethra

o Cervix …………. Vagina

12
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 through
written document & make available at every patient
13
care unit.
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.
14
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).

o CSF: 5 – 10 ml.

15
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. 16

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.

17
Specimen collection…..

6. Stage of the disease at which the specimen is

collected

o Enteric pathogens are present in great numbers

during the acute or diarrheal stage of intestinal
infection.

18
Specimen collection…..
7. Labeling
❖ Make sure that you are collecting/drawing the right
person first.
❖ Then label with:
o Patient name
o Unique identification number
o Patient demographic information
o Specimen collection date
o Specimen collection location
o Diagnostic test results
19
Specimen collection…..

❖ During Labeling:
o Make sure that container label & the requisition match.
o Label should be on the container not on the lid, since the
lid can be mistakenly placed on a different container.
o Ensure the labels on the containers are adherent under
refrigerated conditions.

20
3.3 General Rejection Criteria
1. Unlabelled 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.

o Call the person who collected it for the identification of

the specimen.

o If he is unable to identify the specimen, the ordering

21
physician will be notified.
General Rejection Criteria……

2. Incorrectly labeled (mislabeled) specimens

o Use same criteria as for Unlabelled 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. 22
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.

23
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.

24
Specimen Transportation
❑ Required when:
o 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. 25
Specimen Transportation......

 This transportation is made by using different

preservation methods

▪ Physical

▪ chemical.

- chemical method of preservation is most

common.

26
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)
27
 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

28
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.

29
Specimen Transportation......

❑ Specimen Storage (Prior to Dispatch)

o Urine can be stored at either freezer or refrigerator at

+4 oc.

o Blood samples should be kept at +4 oc.

o Serum & plasma can be stored either frozen or at

+4oc.

30
 Specimen Transportation......

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

dispatch to the laboratory.

❑ Transport

o Urgent requests within normal laboratory hours.

o Transport with labelled, tightly fitted, leak proof

container.

o Wet ice or ice pack should be used.

31
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.

32
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.

33
Stool collection………..

▪ Time of collection

o Collection should always be before:

◼ Radiological studies - barium sulphate for at least

a week.

◼ Certain medication

✓ Anti-protozoa treatment = 3 – 4 weeks after.

✓ Anti-helminithes treatment = 1 - 2 weeks after.

34
✓ Anti-taenia treatment = 5 - 6 weeks after.
Stool collection………..

▪ Preservation of stool

o If possible, process:

✓ Liquid stool: < 30 minutes of passage, RT.

✓ Semi-formed stool: < 1 hour of passage, RT.

✓ Formed stool: < 24 hours of passage, 4 0C.

o If delay is unavoidable, place in suitable preservative

or transport medium
35
3.4.2. collection of Urine

A urine specimen is submitted to the lab in case of

possible renal problems or metabolic disorders of the
body.

 The urine specimen can be referred to urinalysis,

clinical chemistry and bacteriology and Parasitology
labs.

36
Collection of urine….

◼ For urine collected in the lab use a:

◼ Clean conical urine jar

◼ Any clean glass container or bottle

◼ Container should be clean, dry, leak-proof and free

from disinfectants.

37
Collection of urine….

2. Personal hygiene before collection of urine

◼ Women: patients should wash the genital area in all
cases.

 Avoid collecting urine specimens during the

menstrual period.

◼ Men: washing is necessary only for bacteriological

examination.
38
Collection of urine ……….

3. Types of urine collection

◼ First Morning Specimen - a specimen obtained during

the first urination of the day.

◼ Random Specimen - a specimen obtained at any time

during examination.

◼ Postprandial : a specimen obtained 2 hours after

meal.

39
Collection of urine ……….

◼ Clean catch: obtaining of urine directly from blader by

catheterization.
◼ 24- Hour specimen - a specimen obtained within 24
hours.
◼ Mid- stream Specimen - a specimen obtained from the
middle part of the first urine.

40
Collection of ……..
4. Urine preservative and stabilizers
Urine preservatives could be physical or chemical
Methods Advantages Disadvantages

Physical Refrigeration this method prevent bacterial the urine sample at [Link]
growth, and so avoid changes that forshort periodof time, ususally from
occur due to the growth of 6to 8 hours preservation.
bacteria. No chemical is used

Freezing Keeping the urine sample below - destroys formed elements. It is also
200c. this method good for not suitable for physical
preserving bilirubin and examination, because of turbidity.
Urobilinogen. Urine preserved by deep freezing,
will increase its specific gravity

Chemica Boric acid Preserves chemicals and formed Precipitate uric acid
l elements
Sodium Carbonate Preserves porphyries and Interfere with other urine
41
urobilinogen constituents
Chemical Toluene (Till it forms thin layer over Preserves acetone, Flammable
the urine) Reducing Substances,
protein

Thymole ( small crystal 5 Preserves most Can cause false

mm diametre/100ml urine) constituents positives for
proteins.
Chloroform (1 tablet/60 ml Preserves urine Settles to the
urine) aldosterole level bottom of the
urine containers
Formaldehyde (1 drop/30 Preserves formed Interferes with
ml urine) elements glucose
evaluation
HCL (1 drop/15 ml urine) Stabilizes Formed elements
42
steroids, catecola are destroyed,
 3.4.3. 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)

43
Blood collection…….
1. Collection sites:
a. Capillary blood
o Ring/middle finger - adults & children – half way b/n
centre & ball (vascular & fleshy).
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.

44
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 differential
count.

45
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.
46
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.

47
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.

48
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.

49
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, the latter is disposable and is
not popular in developing countries because of the high
cost.

50
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).

51
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).

52
Veins of fore arm

53
Blood collection…….

Equipment and Supplies

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

54
Blood collection…….

55
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, and hemolytic.
56
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.

57
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 veinpuncturist to the patient to minimize
apprehension.

- Never draw blood from standing patient or patient sitting

on a high stool. 58
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.

59
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.
60
 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
61
blood in to appropriate container.
 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.

62
Sites to Avoid for Venipuncture

◼ Scarred areas

◼ Side of the mastectomy

◼ 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 63

3.4 .[Link] 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
64
Sputum…….

◼ Collection Method
 Provide the patient with a sterile wide- mouthed glass
bottle of about 50 ml capacities with a screw top.
 The laboratory technician or the nurse should be
present when the sample is taken.
 First, the patient should be standing, if possible.
 Then, 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.
65
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. 66
Sputum……..

◼ Liquid frothy saliva and secretions from the nose and

pharynx are not acceptable expectations. 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.

67
Sputum…..

68
3.4.5. Throat, eye, ear, Vaginal and
urethral swab collection
Materials required

 Sterile cotton wool

 Applicator stick/aluminum wire (18cm long and 2mm

in diameter)

69
Throat, eye, ear, …

◼ Method
 Prepare strips of cotton wool, 6cm long by 3cm wide
and as thin as possible.

 Roll the cotton wool round one end of the stick. If

metal wire is used, flatten the end first.

 Mould the swab in a conical shape

 Place in a thick Pyrex tube. Plug with non-absorbent

cotton wool sterilize. 70
A. Collection of specimen from the throat

A throat swab is submitted for the diagnosis of

infection in the upper respiratory tract.

 Collect the specimen from the inflamed part of the

throat after opening the mouth wide, use a tongue
depressor to reach the rear part of the mouth.

71
B. Collection of urethral discharge
◼ If possible, take the specimen first thing in the morning
before the patient has urinated. If necessary clean the
meatus with a swab moistened with sterile Imodium
Chloride solution.

◼ Exert a slight pressure on the penis so that a drop of pus

appears at the meatus.

◼ Remove the pus with a sterile inoculating loop or apply

directly to a clean slide.
72
 If no pus appears, insert the sterile loop approx. 25cm
up the urethral canal to attain a specimen.

c. Collection of vaginal discharge

 The specimen should be taken by the physician or

specialist nurse from the cervical canal.

73
3.4.6. Collection of Cerebro-spinal fluid
(CSF)

◼ Where is CSF found?

 CSF is contained in the cavity that surrounds the

brain in the skull and spinal cord in the spinal column.

 It nourishes the tissues of the CNS and helps to

protect the brain and spinal cord from injury.

74
Collection of CSF …

◼ Therefore, considerable risk is involved in

collecting a cerebrospinal fluid (CSF) specimen
and hence it is done only by an attending physician
in the hospital or by an experienced nurse.

◼ Lab. Study of CSF is for the diagnosis of problems

related to the brain or CNS.

75
CSF….

◼ Example N. meningitides is the most commonly sought

bacterial organism in CSF.
 The volume of the CSF in adults is 100-150ml.
Method
 The specimen is collected only by:

A physician or

A specially trained nurse

76
CSF…….
Precautions to be taken when examining the CSF in the
lab
 Do not delay in testing the CSF cells and
trypanosomes are rapidly lysed once the CSF is
removed.
 Glucose too is rapidly destroyed, unless
preserved with fluoride oxalate.

 Work carefully and economically often only a small

quantity of CSF is available for examination.
 The specimen is difficult to collect so do not waste it.

77
3.4.7. SKIN SPECIMEN

Skin specimen is used for diagnosis of

 Bacterial diseases Eg. [Link]

 parasitic diseases . O. volvulus

 Fungal disease

78
3.4.8. Collection and transport of semen
 When doing a semen analysis it is important to be
certain that the patient has been correctly identified
prior to collection.
 At some locations the patient is asked to show a
picture ID at the time of collection or when accepting
a specimen produced at home.

In situations where rape is suspected or paternity is in

question, law enforcement officials should be
consulted.

79
Semen …

◼ It may be necessary for law enforcement officials to be

present to identify the patient and to ensure that chain of
custody has been correctly established.

1. Give the person a clean, dry, leak-proof container, and

request him to collect a specimen of semen at home
following 3–7 days of sexual abstinence.

80
Semen …

◼ Note: When a condom is used to collect the fluid,

this must be well-washed to remove the powder
which coats the rubber.
◼ Coitus interruptus: This method of collection
should not be used because the first portion of the
ejaculate (often containing the highest
concentration of spermatozoa) may be lost.
◼ Also the acid pH of vaginal fluid can affect sperm
motility and the semen may become contaminated
with cells and bacteria.

81
Semen…

2. Ask the person to write his name on the container,

date and time of collection, period of abstinence, and
to deliver the specimen to the laboratory within 1 hour
after collection.
 During transit to the laboratory, the fluid should be
kept as near as possible to body temperature.
 This is best achieved by placing the container inside a
plastic bag and transporting it in a pocket in the
person’s clothing.

82
Summary question

1. Explain how to collect, handle and transfer laboratory

specimens.

83
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 [Link] Laboratory manual for
tropical courtiers, Cambridge Univerity press, 2000
(Vol. I ).
4. Chees brough [Link] 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. [Link] 84
 End of slide

Next: chapter will be Reagents

(solutions) used in Clinical laboratory

85