SAMPLE COLLECTION
METHOD-CSF,BODY
FLUIDS,THROAT SWAB AND
SPUTUM
[Link] KAPOOR
CEREBROSPINAL FLUID
• Label 3 fresh sterile screw capped containers with
patients name, identity no, date and time of collection.
• Wear the PPE i.e. surgical cap and mask ,gown, sterile
gloves that are impermeable to liquids and change gloves
between every patient.
• Put sterile sheet or sterile side of the gloves packet on the
bed under the patients lower(lumbar)spine.
• Ensure that the patient is kept motionless during the
lumbar puncture procedure, either sitting or lying on the
side, with his back arched forwards so that the head
almost touches the knees.
• Clean the lumbar spine area along a line drawn
• between the iliac crests with 70% alcohol to remove
debris and oil.
• Disinfect the skin with povidine-iodine using circular
motion from centre outwards.
• Allow the prepared skin to dry.
• Position sterile spinal needle with stylet between the
2 lumbar vertebrae at level L3-L4, L4-L5,L5-S1 and
introduce into the skin with the bevel facing up, in
the backs midline pointing towards the patients
umblicus.
• When the subarachnoid space is reached remove the
stylet,spinal fluid will appear in the needle hub.
• Measure the hydrostatic pressure with a manometer.
• Collect about 1ml CSF into each sterile pre-labelled
tubes slowly at a rate of 4-5 drops per second.
• Not more than 3-5ml of CSF should be collected.
• Remove the needle and place in sharp puncture
resistant container.
• Cover the insertion site with a sterile adhesive
bandage.
• Remove the PPE and discard in appropriate containers.
• Wash hands with soap and water.
• Transport CSF to microbiology lab within 1 hour for
culture and analysis. If not possible inoculate into T-I
medium. If T-I medium is also not available and delay in
processing is inevitable then store at [Link] not
refrigerate.
• Tube 1 for chemical analysis i.e. protein,glucose.
• Tube 2 for microbiology i.e. gram staining,culture etc.
• Tube 3 for hematology i.e. performing cell
count :WBCs
BODY FLUIDS
• Are obtained from sterile sites by percutaneous
aspiration and include pleural, pericardial, peritoneal,
amniotic and synovial fluids.
• Care should be taken to avoid contamination with
normal flora.
• Clean the needle puncture site with 70% alcohol and
disinfect with an iodine solution i.e. 1-2% tincture of
iodine or a 10% sol of povidine iodine to prevent
specimen contamination.
• Aseptically perform percutaneous aspiration with syringe
and needle to perform the tap. Always wear the PPE.
• Specimen should not be send to the lab with needle still
attached as at can cause needle stick injury to the
handler rather the sample should be collected in a proper
screw capped container.
• The lid and outside of the container should not be soiled
with the sample and only adequate amount of fluid
should be sent for examination rather than the whole
aspirate.
• Do not refrigerate.
• Specimen should be labelled properly with patients
name,id no,date, time, site of collection and probable
diagnosis for proper processing.
• Specimen should be collected prior to antimicrobial
therapy for greater diagnostic senstivity.
• Body fluids should not be collected and send on
swabs as quantity of sample may not be sufficient to
ensure recovery of a small no of organisms.
SPUTUM
• The best time to collect a sputum specimen is upon
awakening in the morning, after a bout of cough as it
contains pooled overnight secretions in which
pathogenic bacteria are more likely to be concentrated.
• Make the collection in a disposable, wide mouthed,
screw capped plastic container of about 100ml capacity.
• Sputum should be collected before beginning any
antibiotic therapy.
• Ask the patient to brush his teeth and gargle with water
immediately before obtaining the specimen to reduce
the number of contaminating oropharyngeal bacteria.
• Instruct the patient to take a deep breath & wait until
he feels material coughed into his throat and then to
spit it directly into the opened container without
spilling over the rim. Tightly screw on the cap of the
container. Wipe off any spilled material on its outside
with a tissue moistened with disinfectant, but take care
not to let any disinfectant enter the container.
• If the patient has difficulty in coughing sputum into the
mouth, postural drainage and appropriate
physiotherapy often cause exudate to move in the
bronchi and stimulate productive coughing.
• Deliver the specimen to the lab as quickly as possible
preferably within 2 hour, for delicate bacterial, viral and
mycoplasma pathogens may die out during any longer
delay.
THROAT SWAB
• Ask the patient to gargle and rinse the mouth.
• Make the patient sit in a comfortable position facing the
light.
• With the patients head tilted back and the throat well
illuminated, depress the tongue with a disposable
spatula so that the back of the throat can be seen.
• Use a plain, albumen coated or charcoal coated cotton
wool swab to collect as much exudate as possible.
• Ask the patient to say “aaa” and rub the swab with
rotation over one tonsillar area i.e. anterior tonsillar
pillar then posterior tonsillar pillar followed by tonsil,
• Then the arch of soft palate and uvula, then similarly
the other tonsil and finally the posterior pharyngeal
wall.
• Replace the swab in its tube with care not to spoil
the rim.
• If the swab cannot be delivered to the lab within
about 1 hr, it should be preferably placed in a
refrigerator at 40C until delivery or preferably it
should be submitted in a tube of transport medium
for bacteriological specimen.
• Throat swabs for virological examination are
collected in the same way but the heads are broken
off into small vials of virus transport medium with
penicillin and gentamycin for virus examination or
with ampicillin for Mycoplasma pneumoniae.