Samplecollection,handlingand

transportation
Muhammad Ilyas
PhD scholar (Biochemistry)

Step 1a : Assemble equipment

for collecting blood:
Gauze pads

Adhesive
bandage

Laboratory sample tubes for blood collection

Blood sampling
systems (Needle
and syringe
system, vacuum
extraction
system with
holder, winged
butterfly system
(vacuum
extraction) or
winged butterfly
system

Tourniquet
(single-use)

Tray for assembling

blood collection
tools

Skin
antiseptic
Rack for
solution:
holding blood
70%
tubes
isopropyl
alcohol Durable marker for writing on
laboratory
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Step 1: Before entering patient room, assemble all equipment

Step 1c : Fill out patient documentation:

Label blood collection tubes with date of collection, patient

name, and his/her
identifier number.
Step 1d : Assemble materials for packaging of samples:
Do

Plastic leak-proof packaging container

NOT
forget to fill out necessary
Disposable (paper) towels

laboratory form

Cooler or cold box, if sample requires refrigeration

For the shipment of samples to the

National Central Laboratory follow
Sample Shipment packaging
requirements (see document "How to
safely ship Emerging and Dangerous
Pathogen sample

Secondary
Container
Tertiary
Container

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Step 2: Put on all personal protective equipment (PPE)

DO NOT ENTER THE PATIENT AREA IF YOU DO NOT HAVE ALL PROTECTIVE GEAR ON

Step 2a: Perform hand hygiene. Duration of the entire procedure: 40-60 sec.

Wet hands with

water and
enough soap to
cover all hand
surfaces.

Step 2b: Put

on a gown.

Right
palm over
left
dorsum
with
interlaced
fingers
and vice
versa,

Rub hands,
palm to
palm,
Back of fingers to
opposing palms
with fingers
interlocked

Palm to
palm
with
fingers
interlace
d

Rinse hands
Rotational
rubbing of left
water.
thumb clasped
in right palm
and vice
Step
2c: versa,
Put on face
protection:

O
R

Quick tips: If the patient has

respiratory symptoms, wear a
medical mask underneath the face
shield.
Step 2d: Put on gloves (over gown cuffs).
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with

Dry hands thoroughly

with single use towel.

Put on a medical mask and

eye protection (e.g. eye
visor/goggles)

Step 3a: Prepare room.

Put infectious waste bags and leak- proof Step 3b: Identify and prepare the patient
and puncture resistant sharps container Introduce yourself to the patient and explain what you will do
into patient room and make sure they are with the blood sample and why.
Make sure that this is the correct patient from whom you wish
ready for use.
Place all blood collection equipment in a to take the blood sample.
place that is easy to access.
Step 3c: Select the site, preferably at the bend of the elbow.
Palpate the area; locate a vein of good size that is visible, straight
and clear.
The vein should be visible without applying a tourniquet
Step 3d. Apply a tourniquet
around
Step
3e: Ask the patient to form a fist so that the veins are more prominent
the arm.
Step 3f: Disinfect the area where you
Tie approximately 45 finger widths
will put the needle.
Use 70% isopropyl alcohol.
Wait 30 seconds for the alcohol to dry.
DO NOT touch the

Step 3g: When using vacuum extraction

system with holder, insert the blood
collector tube into the holder.
Avoid pushing the collector tube past the
recessed line on the needle holder or
you may release the vacuum.

Step 3h: Anchor the vein by holding the patients arm and

BELOW the place where you want to

place the needle.
placing a thumb

DO NOT touch the disinfected site.

DO NOT place a finger over the vein to guide the needle.

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j: When blood starts to flow, ask patient to open his/her Step

hand.3m: Remove blood collector tube from holder and put into rack

Step 3k: Once sufficient blood has been collected

(minimum 5ml), release the tourniquet BEFORE
withdrawing the needle

Step 3n: Put needle into leak-proof and puncture

resistant sharps container.
If the sharps container DOES NOT HAVE a needle
remover:
Put the needle and holder into a sharps container.
Do not remove the needle from the holder.
Do not reuse the needle.

If the sharps container HAS a needle remover:

Step 3o: Stop the

bleeding
and clean the skin.

Remove the needle following instructions on the sharps container.

Put the holder into the infectious waste bag for disinfection.

Step 3l: Withdraw the needle gently.

Give the patient a clean gauze or dry
cotton wool ball to press gently on the
Step 3p: Put items that drip
site.
blood
have
body
fluids on
Ask the patient NOT
to or
bend
the
arm

Do not leave patient until

bleeding
has stopped.
Put an adhesive bandage on
the
site, if necessary.

them into the infectious

waste bag for destruction.

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Type of Waste
Hospital waste can be broadly be defined into 2 categories

Risk Waste

Infectious Waste

Pathological
Waste

Shar
ps

Pharmaceutical Waste

Non Risk Waste

Non risk waste is that which
is comparable to normal
domestic
garbage
and
presents no
risk,
greater
therefore,
than waste from
a normal home i.e.

Chemical Waste
Radioactive Waste

Paper
Packaging
Food Waste
15

Waste Disposal
Treatment:

Incineration
Chemical Disinfection
Autoclaving
Encapsulation
Microwave irradiation
etc.,

Final Disposal

Landfill
Burying inside Premises
Discharge into Sewer etc.,

Waste Minimization
Waste not to be Incinerated

Pressurized gas containers

Large amounts of reactive chemical waste
Radioactive waste
Silver salts or radiographic waste
Halogenated plastics (e.g. PVC)
Mercury or cadmium
Ampoules of heavy metals

10

Colour code for Waste Segregation

Different color coding has to be assigned to
various
Blackwaste
:
waste.
forNon-Risk
effective segregation,
as:
Risk waste with Sharps.
Red:
Risk Waste without sharps.
Blue:

v.
Yellow:

Radioactive waste

Green:
Cadmium

Chemicals like Mercury &

All this segregation should be done by the

individual user.
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Step 4: Prepare blood sample for transport

Step 4a: Take the blood tube from the tray and wipe the
blood tube with a disposable paper towel.

Step 4c: Place all items that came into contact with
blood into the infectious waste bag for destruction.

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Step 4: Prepare blood sample for transport

Step 4d: Protect the sample from breaking during transport

by wrapping the tube of blood in a paper towel.

Step 4e: Ask the designated assistant to approach the

patient room, without entering.
This person should have gloves on.
This person should come close to you holding the open
plastic leak-proof packaging container.
This person should not enter the patient room.

Step 4f: The person who has collected the blood sample
should put the wrapped tube of blood into the plastic
leak- proof packaging container.

Be careful not to touch outside of leak-proof

plastic tube with gloves.

Labeling specimens for transportation

Patients name
Clinical specimen
Unique ID number
(Research/Outbreak)
Specimen type
Date, time and place of
collection
Name/ initials of collector

Blood for smears

Collection
Capillary blood from finger prick

make smear
fix with methanol or other
fixative
Handling and transport
Transport slides within 24 hours
Do not refrigerate (can alter cell
morphology)

Blood for cultures

Collection
Venous blood
infants: 0.5 2 ml
children: 2 5 ml
adults: 5 10 ml
Requires aseptic technique
Collect within 10 minutes of fever
if suspect bacterial endocarditis:
3 sets of blood culture

Blood for cultures

Handling and Transport

Collect into bottles with infusion broth

change needle to inoculate the

broth
Transport upright with cushion

prevents hemolysis
Wrap tubes with absorbent cotton
Travel at ambient temperature
Store at 4oC if cant reach laboratory in 24h

Serum

Collection
Venous blood in sterile test tube

let clot for 30 minutes at ambient

temperature
glass better than plastic
Handling
Place at 4-8oC for clot retraction for at least 12 hours
Centrifuge at 1 500 RPM for 5-10 min

separates serum from the clot

Serum

Transport
4-8oC if transport lasts less than 10 days
Freeze at -20oC if storage for weeks or
months before processing and shipment
to reference laboratory
Avoid repeated freeze-thaw cycles

destroys IgM
To avoid hemolysis: do not freeze
unseparated blood

Cerebrospinal fluid (CSF)

Collection

Lumbar puncture
Sterile tubes
Aseptic conditions

Trained person

CSF
Handling and transportation

Bacteria
preferably in trans-isolate medium,
pre-warmed to 25-37C before
inoculation
OR

transport at ambient temperature (relevant

pathogens do not survive at low
temperatures)

Viruses
transport at 4-8oC (if up to 48hrs or -70oC for
longer duration)

Stool samples

Collection:
Freshly passed stool samples

avoid specimens from a bed pan

Use sterile or clean container

do not clean with disinfectant

During an outbreak - collect from 10-20 patients

Rectal swabs

Advantage
convenient
adapted to small children, debilitated patients
and other situations where voided stool
sample not feasible
Drawbacks
no macroscopic assessment possible
less material available
not recommended for viruses

Stool samples for viruses

Timing
within 48 hours of onset
Sample amount
5-10 ml fresh stool from patients (and controls)
Methods
fresh stool unmixed with urine in clean, dry and sterile container
Storage
refrigerate at 4oC; do not freeze
store at -15oC - for Ag detection,polymerase chain reaction (PCR)
Transport
4oC (do not freeze); dry ice for (Ag detection and PCR)

Stool samples for bacteria

Timing
during active phase
Sample amount and size
fresh sample and two swabs from patients,
controls and carriers (if indicated)
Method
Cary-Blair medium
For Ag detection/PCR no transport medium
Storage
refrigerate at 4oC if testing within 48 hours, -70oC if
longer; store at -15oC for Ag detection and PCR
Transport
4oC (do not freeze); dry ice for Ag, PCR detection

Stool samples for parasites

Timing
as soon as possible after onset
Sample amount and size
at least 3 x 5-10 ml fresh stool from patients and
controls
Method
mix with 10% formalin or polyvinyl chloride, 3 parts stool
to 1 part preservative
unpreserved samples for Ag detection and PCR
Storage
refrigerate at 4oC; store at -15oC for Ag detection and
PCR
Transport
4oC (do not freeze); dry ice for antigen detection and PCR

Throat swab
(posterior pharyngeal swab)

Hold tongue away with

tongue depressor
Locate areas of
inflammation and exudate
in posterior pharynx,
tonsillar region of throat
behind uvula
Avoid swabbing soft
palate; do not touch
tongue
Rub area back and forth
with cotton or Dacron
swab

WHO/CDS/EPR/ARO/2006.1

Nasopharyngeal swab

Tilt head backwards

Insert flexible fine-shafted
polyester swab into nostril
and back to nasopharynx
Leave in place a few seconds
Withdraw slowly; rotating
motion

WHO/CDS/EPR/ARO/2006.1

Naso-pharyngeal aspirate

Tilt head slightly backward

Instill 1-1.5 ml of VTM /sterile normal
saline into one nostril
Use aspiration trap
Insert silicon catheter in nostril and
aspirate the secretion gently by
suction in each nostril

WHO/CDS/EPR/ARO/2006.1

Sputum

Collection
Instruct patient to take a deep breath and
cough up sputum directly into a wide-mouth
sterile container
avoid saliva or postnasal discharge
1 ml minimum volume

Step 5: Remove Personal Protective Equipment (PPE)

Step 5a: Remove the gloves.

Step 5b: Remove the gown

1.Grasp the outer edge of the 1st glove and peel

it off.

2.Hold the 1st glove in 3. Remove 2nd glove from

the gloved hand and drag
the inside, creating
a
a bare finger under the "bag for both gloves and 2
glove. throw it in waste
bag for
disposal.

Untie the gown

nd

p 5c: Perform Hand hygiene. Duration of the entire procedure: 40-60 sec.
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Step 5d: Take off face protection

Remove face shield from behind.
If it is a reusable face shield, place it in an
infectious waste bag
for disinfection.
If it is a disposable face shield, place it in an
infectious waste bag for destruction.
Optional: If wearing a medical mask, remove the
medical mask from behind, starting with the
bottom strap, and place it in a infectious waste
When
wearing goggles
bag for destruction.

and a mask:

Remove goggles from behind.

If reusable goggles, place it in
an infectious waste bag for
disinfection.
If disposable goggles, place it
in an infectious waste bag for
destruction.
Remove the medical mask
from behind, starting with the
bottom strap, and place it in

5e: Perform Hand hygiene. Duration of the entire procedure: 40-60 sec.

Quick Tips:
Place all reusable equipment into a separate infectious waste
bag for disinfection.
When collecting blood samples from multiple patients:
Change gloves between each patient.
Wash hands between each patient.
DO NOT WASH GLOVED HANDS.
DO NOT REUSE GLOVES.