SPECIMEN COLLECTION,

HANDLING ,AND PROCESSING FOR

MICROBIOLOGY TESTING

GENERAL CONCEPTS FOR SPECIMEN

COLLECTION, HANDLING AND PROCESSING

Quality of the clinical specimen is extremely

important
Careful and appropriate collection techniques
must be observed
Transport to the laboratory must be timely, avoid
delay
The clinical specimen must be processed
immediately, avoid delay

GENERAL GUIDELINES
Collect specimen prior to administration of
antibiotics
Timing is important
Collect adequate amount of sample
Know the criteria for acceptance and rejection of
clinical specimens
Evaluate whether the agent isolated and
identified would point out to an infectious process
or merely points out to contamination

SPECIMEN COLLECTION, HANDLING AND

PROCESSING FOR BACTERIOLOGIC TESTING

Clinical Specimen
1. Urine
2. Stool
3. Sputum and lower resp. tract (LRT) samples
4. Throat and upper resp. tract (URT) samples
5. CSF
6. Blood
7. Skin, wound and tissue
8. Genital discharge

1. URINE CULTURE
For diagnosis of UTI
Collection techniques:

1. midstream, clean-catch
2. catheter
3. SPA
Pointers: Use sterile container
Cleanse area first with soap and water
before sample is collected; for SPA disinfect skin
first

URINE CULTURE
Transport: within 24hrs at 40C
Storage before processing: 24 hrs at 40C
Plating media: BAP, MacConkey agar
(+Anaerobic agar for SPA)
: Perform colony count
(plate quantitatively on BA)
Direct examination: GS

2. STOOL CULTURE
For diagnosis of GIT bacterial infections
Collection techniques:
1. direct
2. rectal swab
Pointers: Use sterile container
Use transport medium when delay in
processing is expected
An enrichment media is used (GN
broth or Selenite broth)

STOOL CULTURE
Transport: 24hrs at 40C use transport media
Storage before processing: 72hrs at 40C
Plating media: BAP, Campy BA, CCF, CIN,
MacConkey or EMB, HE ,SSA, XLD, TCBS
Direct examination: MB for fecal leukocytes

3. SPUTUM AND LOWER RESP. TRACT

CULTURES
For diagnosis of LRTIs
Collection technique:
1. deep cough method in AM
2. aspiration of gastric
contents (for children)
3. BAL (less invasive)
4. TTA, thin needle
aspiration, open-lung biopsy, thoracentesis
(pleural fluid)
5. induced sputum collection

SPUTUM AND LOWER RESP. TRACT

CULTURES

Pointers: Use sterile container

Choice of method for collection must be
considered
Collect specimen with minimum or less
contamination
Evaluate the quality of specimen

SPUTUM AND LOWER RESP. TRACT

CULTURES
Transport: 24hrs at RT
Storage before processing: 24hrs at 40C
Plating media: BAP, CAP, MacConkey
Direct examination: GS, AFB or other special
stains

4. THROAT AND UPPER RESP. TRACT

CULTURES
For diagnosis of URTIs
Collection technique:
1. Throat swab (pharynx)
2. Nasopharyngeal swab
Pointers: Use sterile container
Ensure proper swabbing of collection
area
Use transport medium (Amies or
Stuarts)

Link:resp_sx

THROAT AND UPPER RESP. TRACT

CULTURES
Transport: 24 hrs at RT
Storage before processing: 24hrs at 40C
Plating media: BAP, CAP, CTA, Loefflers agar
slant, MTM agar
Direct examination: use antigen testing

5. CSF
For diagnosis of CNS infections
Collection technique:
1. Lumbar puncture
2. Collection from shunt

Pointers: Use sterile container, use aseptic

technique
Specimen must be processed
IMMEDIATELY, DO NOT DELAY

CSF
Transport: immediate at RT
Storage before processing: 6hrs at 370C, for viral
culture 3 days at 40C
Plating media: BAP, CAP, MacConkey, Anaerobic
agar
Direct examination: GS, MB, AO, AFB,rapid
tests

6. BLOOD CULTURE

For diagnosis of bloodstream infections (BSIs)

Collection technique:
1. Venipuncture
Pointers: Use sterile container
Use aseptic technique for collection
Timing of blood collection is IMPORTANT
Number of collection (# of set) is considered
Tubes or bottles contain SPS (1 aerobic, 1
anaerobic) ; other culture media include: SCD or TSB,
Columbia broth, thioglycollate broth, BHIB,
supplemented peptone broth (SPB)

BLOOD CULTURE
Transport: 2hrs at RT
Storage before processing: 370C incubated
Plating media: BAP, CAP, MacConkey, Anaerobic
agar, BHI agar
Direct examination: GS, Giemsa, MB, AO, AFB

SKIN, WOUND AND SOFT TISSUE

Transport:24hrs at RT; moisten swab with TM
Storage before processing: 24 hrs at RT
Plating media: BA,CA, MacConkey

URO-GENITAL TRACT SPECIMEN

Transport: 24 hrs at RT
Storage before processing: 24 hrs at RT
Plating media: BA, CA, TM