COLLECTION AND TRANSPORTATION OF CLINICAL SAMPLES

Principles of Sample Collection:

Aseptic precautions to minimize chances of
contamination.
Appropriate anatomic sites
Adequate volume
Adequate no. of samples
Appropriate time
Appropriate container with proper labelling
Before initiation of anti-microbials
Adequate information in request form

1. COLLECTION AND
TRANSPORT
OF CLINICAL
SAMPLES

2. Principles of Sample Collection:
Aseptic precautions to minimize chances of
contamination.
Appropriate anatomic sites
Adequate volume
Adequate no. of samples
Appropriate time
Appropriate container with proper labelling
Before initiation of anti-microbials
Adequate information in request form

3. 1.Blood
2.Body fluids
Cerebrospinal fluid
Pleural fluid
Peritoneal fluid
Pericardial fluid
Synovial fluid
Amniotic fluid
Broncho-alveolar lavage
3.Urine
4.Bone marrow
Samples that are normally sterile

4. BLOOD

5. Collection of Blood
•Serology
-collect blood in plain vial / vaccutainer
•Blood Culture
-collect blood in blood culture bottle
(Brain Heart Infusion Broth/
BacT/Alert bottles)

6. Blood Collection-Steps
-Patients’ blood
must be collected
byaseptic
venepuncture.

7. .
-Once a vein is
selected, skin site
is disinfectedwith
70% isopropyl
alcohol, in a circle
approx 5cm in
diameter.
-Allow it to air dry.
Blood Culture

8. -Insert the needle into
the vein and
withdraw blood.
-5 to10 ml of bloodis
inoculated through
the lid, into the
blood culture bottle
(without opening the
lid).

9. AEROBIC
BacT/ALERT
PEDIATRIC
BacT/ALERT
ANAEROBIC
BacT/ALERT
8 -10ml 0.5 -4ml 8 -10ml
Routine evaluation for
sepsis should
Include submission of TWO
aerobic blood cultures
collected from different
venipuncture sites or from
the same site 15 min apart
Used instead of the
aerobic bottle in
patients from whom 8 -
10ml blood cannot be
obtained
MUST be used in
conjunction with the
aerobic bottle and should
be used ONLY on patients
at risk for sepsis with
anaerobic organisms

10. Blood Culture sample
Acute sepsis-2-3 sets of blood cultures
from separate sites
Endocarditis-3 cultures over 1-2 hours, 15
minutes apart (repeat if negative after 24
hours)
PUO-2 cultures, at least 1 hour apart (if
negative -repeat 2 cultures 1 hour apart,
after 24-36 hours)

11. URINE

12. URINE SAMPLE COLLECTION &
TRANSPORT
Mid stream clean voided urine sample ( most common)
Catheter sample
Suprapubic aspiration
-Sample collected in sterile wide mouthed leak proof &
disposable container
-Sent to laboratory within ½ hr of collection
-Sample refrigerated at 4°C in case of delay
-Addition of 1% boric acid to sample if refrigeration not
available

13. Mid stream sample collection
-Area around urethral meatus cleaned with sterile saline
soaked gauze piece. No antiseptics to be used.
-Min 20 ml sample required for culture

14. Supra pubic aspiration
from bladder
-Ideal sample in case of UTI
-Sample sent as closed syringe
technique for anaerobic culture

15. Catheter sample collection
Two methods:
1.Collection using sterile 26 SWG needle and syringe at
junction of catheter tubing and urine bag
2. Clamping of catheter at junction with urine bag for 15
min to allow urine collection. Disconnection of junction
with collection of urine sample into wide sterile test tube.
No sample to be taken from urine bag

16. For fungal culture-
-Early morning first voided urine –2 consecutive days
For TB culture-
-Multiple (5) samples of first passed early morning urine.
-24 hrs urine collection not recommended to avoid dilution
of mycobacteria by contaminants from progressive
samples

17. BODY FLUIDS

18. Pleural fluid
-Sample collection through aseptic
technique after skin disinfection
preferably imaging guided
-Fluid collected in at least two sterile test
tubes
-Additional tubes for cytology &
biochemistry
-If pure pus aspirated from pleural cavity
(empyema thoracis), atleast 5 ml
sample should be transported
immediately to lab using closed
syringe technique(for anaerobic
culture)

19. Peritoneal fluid
-Aspirated using aseptic technique after skin
disinfection and collected in sterile tubes
-Amount of fluid req:
5-7ml –aerobic culture
5ml-anaerobic culture in anaerobic vial
10ml-for fungal culture
-Immediate transport to lab in c/o suspected
gonoccocal or non sporing anaerobic
infections
>40ml fluid or whole bag-For Continuous
Ambulatory Peritoneal Dialysis

20. CSF
•Sample collected by lumbar
puncture using aseptic technique
•Collected in 3-4 tubes
•Initial portion to be collected in
sterile tubes 1 & 2:-sent for C/S
•Tubes 3 & 4:-sent for clinical
pathology
•Sample sent to laboratory as
early as possible
•Sample should not be
refrigertaed as organisms such as
meningococcus & pneumococus
get killed
•Kept in incubator / at room
temperature in case of delay

21. Synovial fluid
Collect in 3 containers:
-Aerobic blood culture bottle
-Anaerobic culture bottle
-Fungal culture bottle
Rest of the fluid should be sent in a sterile tube for cytology
and biochemistry

22. RESPIRATORY
TRACT

23. UPPER RESPIRATORY TRACT

24. Throat Swab
Technique-
-Ask the patient to say “aah”
-Press the tongue down with a tongue depressor
-Adequate source of light required
-Sterile swab rubbed against tonsils, uvula &
posterior pharyngeal wall
-Swab removed from mouth without touching sides
of oral cavity.
-At least 2-3 swabs should be collected
Transported within 2 hrs of collection
If delay, transported in Stuart’s medium

26. Anterior nasal swab
-To detect carriers of C.diphtheriae
& MRSA
-Sterile cotton swab rubbed against inner sides of nasal
mucosa
Pernasal swabs
-To detect B.pertussis
-Sterile cotton or calcium alginate swab attached to bent
piece of wire is passed along floor of nostril, downwards &
backwards as far as nasopharynx
Nasopharyngeal Aspiration
-In children-when it is not possible to obtain sputum

27. LOWER RESPIRATORY
TRACT SECRETIONS

28. Sputum
Tracheobronchial Secretions:
-Collected during acute stage of
disease
-Early morning deep cough
-Assisted sputum production by
physiotherapy
For TB:
3 or more early morning samples
For fungal infections:
24-hour collection over 3 days
Transported within 1 hour ; if
delay-refrigerate at 4
0
C

29. Bronchoscopy:
Used for-
1. Broncheolo-alveolar Lavage (BAL)
-Most important representative of all LRT samples
-Transported immediately; if delay –keep at 4
0
C
2. Lung biopsy

30. BAL FLUID

31. Pus/Wounds/Burns

32. Abscesses-drained using needle & syringe; sent directly to
lab
For anaerobic cultures-sent as closed syringe technique
within 10 min
If delay of > 1 hr-incubate at 37
0
C
No pus sample should be refrigerated

33. Wound Swab
-Indicated : when there is no discharge from wound area
-Collected from central necrotic area
for aerobic & anaerobic cultures
-For aerobic culture swab transported in Amies Transport
medium
-For anaerobic cultureswab transported in Stuart’s medium
–swab stabbed into upper 1/3 of medium with its stick
broken off and the tube sealed. Sent to lab within 10min
of collection.

34. STOOL

35. -Sample to be collected in clean,
dry, disinfectant-free container
Rectal swab:
Indications:
-stool sample collection not possible
-rectal ulcers
cotton wool swab inserted into
rectum for 10 sec,avoiding
contamination from perianal area
Transport media:
Cary-Blair medium-Salmonella,
Shigella,Vibrio,Yersinia (48 hours)
& Campylobacter (6hours)

36. SAMPLES THAT SHOULD
BE REFRIGERATED IN
CASE OF DELAY IN LAB
TRANSPORTATION
-URINE
-SPUTUM
-FAECES
-NASOPHARYNGEAL
SAMPLES FOR CULTURE
-ALL GUT SAMPLES FOR
CHLAMYDIAE
-ALL CATHETER TIPS

37. SAMPLES THAT
SHOULD NOT TO
BE REFIGERATED
-BLOOD
-BODY FLUIDS
-PUS SAMPLES
-SAMPLE SUSPECTED
TO CONTAIN NISSERIAE
& HEMOPHILUS
-WOUND SWABS