RESPIRATORY SPECIMEN COLLECTION
Royal Center for Disease Control
Supplies Needed
for Specimen Collection and Transport
 Viral Transfer Medium (VTM) with sterile swabs in zip-locked bag
 Dry ice or Ice or Ice packs (temporary storage and specimen transfer)
 Cold box / Styrofoam box
 Pre-printed barcode labels
 Glove, Mask and Lab gowns
 Tongue depressor (optional)

Rapid Diagnostic Test Kit
Specimen Collection
Polyester Fiber-Tipped Swab
 Should be synthetic material
(e.g. drayon, rayon, or
polyester-fiber swabs)
 Do not use calcium alginated or
cotton swabs nor ones with
wooden sticks; they inhibit
certain tests (e.g. PCR)
Specimen Collection
Viral Transport Medium (VTM)
• Used in the collection of samples
for viral isolation and PCR testing
• Prevents specimen from drying out
• Prevents bacteria and fungi growth
• Can be home-made or commercial
• Do not use if cloudy in appearance
or change in color
Respiratory Specimen Collection
You will most commonly collect:
 Nasal swab
 Throat (Oropharyngeal) swab
Specimen for Laboratory Tests
Throat swab or Nasal swab are technically simple to perform
under field conditions.
Up to 3 swabs will be collected from one patient:
 Two swabs to be put in a single Viral Transport-Medium (VTM)
tube for shipment to AFRIMS laboratory
AND
 One nasal swab for on-site Rapid Test (Use the swab that comes with
the Rapid Test Kit)
2 throat swabs
1 nasal swab for Rapid test
Throat Swab Procedure
• Vigorously rub the tonsils,
then the posterior pharynx
while rotating the swab to
collect the specimen on all
sides of the swab
• Two throat swabs can be
performed simultaneously
Labeling the VTM Tube
• Place the swabs into the VTM tube,
and break off the tip of the swabs (do
not break it too short) so that the top
can be closed securely leaving the
swabs in place
• AFRIMS will send you pre-printed
labels with specimen numbers to affix
to the VTM tubes. Please do not write
patient names on the label
Pre-printed Labels
FLU-Bhutan (WRAIR#1599)
Site Code
Specimen No.
Study site
A
FLU-BTA-00000
B
FLU-BTB-00000
Eastern Regional Referral Hospital, Mongar
Jigme Dorji Wanghck National Referral Hospital,
Thimphu
C
FLU-BTC-00000
Paro Hospital
D
FLU-BTD-00000
Punakha Hospital
E
FLU-BTE-00000
Trongsa Hospital
F
FLU-BTF-00000
Phuentsholing Hospital
G
FLU-BTG-00000
Central Regional Referral Hospital, Gelephu
H
FLU-BTH-00000
Trashigang Hospital
I
FLU-BTI-00000
Damphu Hospital, Tsirang
Nasal Swab Procedure
• Gently insert the swab
into one nostril and rub
while rotating the swab
from a depth of 2-3
centimeters by using the
swab provided with the
rapid diagnostic test kit
Rapid Diagnostic Test
Rapid Test - Basic Facts
• The test contains four different antibodies:
two for detection of influenza A and two for
detection of influenza B
• It can give a positive result in 2 to 10 minutes,
depending upon the level of virus present in
the specimen
• The kit is stable for 24 months at room
temperature (15º to 30ºC)
Rapid Diagnostic Test
QuickVue Influenza A+B test (Quidel)Immunochromatography
Sterile nasal swabs in
individual pouches
Extraction tubes
Extraction reagent solution
Package insert
and Procedure card
Positive influenza type A, B
and Negative control swab
Individual packaged strips
Rapid Test Procedure
Consult kit package insert for full instructions for use.
Interpretation of Results
Rapid Test
•
Advantages
–
–
–
–
•
Fast
Relatively simple
Can be done on-site
May be useful to guide clinical care
Disadvantages
– Cannot subtype beyond flu A and B (i.e. cannot distinguish
pandemic subtypes from seasonal flu)
– Less sensitive than laboratory methods
– Low sensitivity/specificity for H5N1
– Unknown sensitivity/specificity for pandemic H1N1
(probably somewhat less sensitive than seasonal flu)
Rapid Test
• Reported sensitivity of about 70-75% and
specificity of 90-99% for seasonal influenza
when compared to virus culture
– BUT wide range of sensitivities reported in medical literature from low to
moderate
• BOTTOM LINE: Negative rapid test does NOT
rule out influenza and clinical judgment
should guide medical care
STORAGE/PACKING/TRANSPORTATION
Specimen Storage
For specimens in VTM:
 Ship specimens to laboratory as soon as possible
 Store specimens at 2-8 °C before and during
shipment (within 72 hours)
 Store specimens at -70 °C beyond 72 hours
 Do not store in standard freezer, especially “frost
free” freezer – keep on ice or in refrigerator instead
Specimen Packing and Transportation
•
–
–
•
–
–
2 main goals
Protect environment and carrier
Protect the sample
International Air Transport Association (IATA)
www.iata.org
Provides guidelines for packaging and shipping by
air
– Categorization of Infectious Substances as
“Dangerous Goods”
Packing Respiratory Samples
for Transportation
• Use 3 packaging layers
• Primary receptacle holds biological material
– i.e. VTM tube wrapped with parafilm
• Secondary container
– Durable, watertight, leak-proof
– Several primary receptacles can go into secondary container
– Place absorbent, cushioned material between primary and secondary
containers
• Outer container
–
–
–
–
Rigid, durable, insulated
Place itemized list of contents between secondary and outer containers
Place ice/cold packs between secondary and outer containers
Outer container should be properly marked and labeled
Packing Respiratory Samples
for Transportation (1)
For transferring sample from “Study Site to the Lab”
• Seal VTM tube with parafilm
• Place VTM within its own
watertight ziplock bag made
for biological specimens
“this is primary receptacle”
Packing Respiratory Samples
for Transportation (2)
Place up to 10 single VTMcontaining bags within
another watertight
container (e.g. biohazard
bag or sturdy plastic
container with lid) along
with absorbent material
“this is secondary
container”
Packing Respiratory Samples
for Transportation (3)
Put secondary
container into a rigid
insulated container
(e.g. styrofoam box)
with ice/cold packs –
this is outer container
Specimens Transportation
• Store specimens at
2-8
°C and send to Laboratory
center (PHL) within 72
hours for storing
specimens at -70 °C
• Specimen can be shipped
by ice or icepack if dry-ice
is not available.
•Specimens
•Cold-chain maintenance table
form
• Store specimens at
-70°C
• Badge all specimens
from all study sites and
ship specimens to
AFRIMS Virology on
dry ice.
•Specimens
•Demographic/Clinical Form
•Cold-chain form
•Consent form
•Enrollment log
• Receive specimens and
perform the laboratory
testing (PCR testing)
BIOSAFETY
STANDARD PRECAUTIONS
 Wash hands with soap as a cost effective control transmission of
infectious by hand
 Wash hands immediately with
running water for at least 20 seconds
after gloves are removed
 Wash hands immediately with running water
for at least 15 minutes in case of hand
exposure to infectious materials
STANDARD PRECAUTIONS
Wash Hands :
 before and after subject/patient contact
 after taking samples
 after removing gloves
 before eating
 before leaving the laboratory working
areas
Personal Protective Equipment (PPE)
 Ensure appropriate personal protective equipment (PPE) is
available
 Wear PPE at all times (Mask, Gloves, Lab coat, Goggle or face
shield)
 Properly use of PPE
BIOSAFETY FOR LABORATORY
 Treat every specimen as if it were an infectious material
 Do not eat, drink, smoke, handle contact lenses, apply
cosmetics and apply medicine in the area which respiratory
specimens are handled
 Do not store food, drink, cigarettes, contact lenses, cosmetics
and medicine for human consumption in the area which
respiratory specimens are handled
SPILL
Materials for a biological spill kit:
• Disposable Gloves
• Rubber Gloves (utility)
• Paper towels
• Forceps or big tweezers
• Bleach
• Mixing / Spray bottle
• Biohazard bags
• Disposable mask, Goggles
or face shield
Instruction for Biohazard Spill
Wear Gloves
 Place an absorbent towel or gauze on the spill
Liberally add a 10% bleach solution to the towel
 Leave the area for at least a half an hour
 Remove the towel and discard in a biohazard bag
 Apply 70% ethanol to the area 2-3 times, wiping the
area thoroughly

WASTES
• Waste Disposal
- Biohazard containers for disposal of contaminated materials
- Double bag system for transport