1 Specimen Collection For Culture

LECTURE NOTES
For Medical Laboratory Students

Specimen Collection For Culture

Dr.Abdulrazzaq Othman Alagbare

MD. Clinical pathologist
Yemeni Jordanian University
Yemen- Sana'a
2016

Prepared by. DR. Abdulrazzaq Othman Alagbare

 2 Specimen Collection For Culture

Specimen Collection For Culture

Blood Culture
Blood, routine (blood culture Whole blood 10 mL in each of 2 bottles for adults and children; 5 mL, if
for aerobes, anaerobes, and possible, in each of 2 bottles for infants; less for
yeasts) neonates
Blood for fungi/Mycobacterium Whole blood 10 mL in each of 2 bottles, as for routine blood cultures, Same as for routine blood culture
spp. or in Isolator tube requested from laboratory
Blood, Isolator (lysis Whole blood 10 mL Isolator tubes
centrifugation)

RESPIRATORY TRACT
ose Swab from nares 1 swab Sterile container or similar Swabs made of calcium alginate may be used.
transport system containing
holding medium
Throat Swab of posterior pharynx, 1 swab Sterile container or similar swab
ulcerations, or areas of specimen collection system
suspected purulence containing holding medium
Sputum Fresh sputum (not saliva) 2 mL Commercially available sputum Cause for rejection: Care must be taken to ensure
collection system or similar that the specimen is sputum and not saliva.
sterile container with screw cap Examination of Gram's stain, with number of
epithelial cells and PMNs noted, can be an
important part of the evaluation process.
Induced sputum specimens should not be
rejected.
Bronchial aspirates Transtracheal aspirate, 1 mL of Sterile aspirate or bronchoscopy Special precautions may be required, depending
bronchoscopy specimen, aspirate or tube, bronchoscopy brush in a on diagnostic considerations (e.g.,
or bronchial aspirate brush in separate sterile container Pneumocystis).
transport
medium

Prepared by. DR. Abdulrazzaq Othman Alagbare

 3 Specimen Collection For Culture

Stool Sp. Collection

Stool for routine culture; stool Rectal swab or 1 g of stool Plastic-coated cardboard cup or If Vibrio spp. are suspected, the
for Salmonella, Shigella, and (preferably) fresh, or 2 rectal plastic cup with tight-fitting laboratory must be notified, and
Campylobacter randomly collected swabs lid. Other leak-proof appropriate collection/transport methods
stool containers are also acceptable. should be used.
Stool for Yersinia, E. coli O157 Fresh, randomly 1g Plastic-coated cardboard cup or Limitations: Procedure requires
collected stool plastic cup with tight-fitting enrichment techniques.
lid
Stool for Aeromonas and Fresh, randomly 1g Plastic-coated cardboard cup or Limitations: Stool should not be cultured
Plesiomonas collected stool plastic cup with tight-fitting for these organisms unless also cultured
lid for other enteric pathogens.
Stool for Yersinia, E. coli O157 Fresh, randomly 1g Plastic-coated cardboard cup or Limitations: Procedure requires
collected stool plastic cup with tight-fitting enrichment techniques.
lid
Stool for Aeromonas and Fresh, randomly 1g Plastic-coated cardboard cup or Limitations: Stool should not be cultured
Plesiomonas collected stool plastic cup with tight-fitting for these organisms unless also cultured
lid for other enteric pathogens.

UROGENITAL TRACT
Urine Clean-voided urine specimen 0.5 mL Sterile, leak-proof container with
or urine collected by screw cap or special urine
catheter transfer tube
Urogenital secretions Vaginal or urethral 1 swab or 0.5 Trans-swab containing Amies Vaginal swab samples for "routine
secretions, cervical swabs, mL of fluid transport medium or similar culture" should be discouraged
uterine fluid, prostatic system containing holding whenever possible unless a
fluid, etc. medium for Neisseria particular pathogen is suspected.
gonorrhoeae; modified Todd- For detection of multiple organisms
Hewitt broth for group B (e.g., group B Streptococcus,
Streptococcus surveillance Trichomonas, Chlamydia, or
cultures Candida spp.), 1 swab per test
should be obtained.

Prepared by. DR. Abdulrazzaq Othman Alagbare

 4 Specimen Collection For Culture
BODY FLUIDS, ASPIRATES, AND TISSUES
Cerebrospinal fluid (lumbar Spinal fluid
puncture)
Body fluids Aseptically aspirated body fluids 1 mL for routine
Biopsy and aspirated Tissue removed at surgery,
materials bone, anticoagulated bone
marrow, biopsy samples, or
other specimens from normally
sterile areas
Wounds Purulent material or abscess
contents obtained from wound
or abscess without
contamination by normal
microflora
Prepared by. DR. Abdulrazzaq Othman Alagbare
1 mL for routine Sterile tube with tight-fitting
cultures; ≥5 mL for cap
Mycobacterium
Sterile tube with tight-fitting
cultures cap. Specimen may be left in
syringe used for collection if
the syringe is capped before
transport.
1 mL of fluid or a 1-g Sterile " container "-type swab
piece of tissue or similar transport system
containing holding medium.
Sterile bottle or jar should be
used for tissue specimens.
2 swabs or 0.5 mL of Culture swab or similar
aspirated pus transport system or sterile
tube with tight-fitting screw
cap. For simultaneous
anaerobic cultures, send
specimen in anaerobic
transport device or closed
syringe.
Do not refrigerate; transfer
to laboratory as soon as
possible.
For some body fluids (e.g.,
peritoneal lavage samples),
increased volumes are
helpful for isolation of
small numbers of bacteria.
Accurate identification of
specimen and source is
critical. Enough tissue
should be collected for
both microbiologic and
histopathologic
evaluations.
Collection: Abscess contents
or other fluids should be
collected in a syringe (see
above) when possible to
provide an adequate
sample volume and an
anaerobic environment.
 5 Specimen Collection For Culture

SPECIAL RECOMMENDATIONS

ungi Specimen types listed 1 mL or as specified Sterile, leak-proof Collection: Specimen should be
above may be used. above for individual container with tight- transported to microbiology laboratory
When urine or listing of specimens. fitting cap within 1 h of collection. Contamination
sputum is cultured Large volumes may with normal flora from skin, rectum,
for fungi, a first be useful for urinary vaginal tract, or other body surfaces
morning specimen is fungi. should be avoided.
usually preferred.
Mycobacterium (acid-fast bacilli) Sputum, tissue, urine, 10 mL of fluid or Sterile container with Detection of Mycobacterium spp. is
body fluids small piece of tight-fitting cap improved by use of concentration
tissue. Swabs techniques. Smears and cultures of
should not be used. pleural, peritoneal, and pericardial
fluids often have low yields. Multiple
cultures from the same patient are
encouraged. Culturing in liquid media
shortens the time to detection.

Prepared by. DR. Abdulrazzaq Othman Alagbare

 6 Specimen Collection For Culture

Legionella Pleural fluid, lung 1 mL of fluid; any size

biopsy, tissue sample,
bronchoalveolar lavage although a 0.5-g
fluid, bronchial/ sample should be
transbronchial biopsy. obtained when
Rapid transport to possible
laboratory is critical.
Anaerobic organisms Aspirated specimens 1 mL of aspirated An appropriate Specimens cultured for obligate
from abscesses or fluid or 2 swabs anaerobic transport anaerobes should be cultured for
body fluids device is required. facultative bacteria as well.
Viruses Respiratory secretions, 1 mL of fluid, 1 swab, Fluid or stool samples Most samples for culture are transported
wash aspirates from or 1 g of stool in in sterile containers in holding medium containing
respiratory tract, nasal each appropriate or swab samples in antibiotics to prevent bacterial
swabs, blood samples transport medium viral culture devices overgrowth and viral inactivation.
(including buffy coats), (kept on ice but not Many specimens should be kept cool
vaginal and rectal frozen) are generally but not frozen, provided they are
swabs, swab suitable. Plasma transported promptly to the laboratory.
specimens from samples and buffy Procedures and transport media vary
suspicious skin lesions, coats in sterile with the agent to be cultured and the
stool samples (in some collection tubes durat
cases) should be kept at 4 to
8∞C. If specimens are
to be shipped or kept
for a long time,
freezing at 80∞C is
usually adequate.

Prepared by. DR. Abdulrazzaq Othman Alagbare

 7 Specimen Collection For Culture

Urine Tests Requiring No Preservatives

Test name is listed with preferred specimen (timed or random collection)

Aldosterone, refrigerate during collection or, 10 g boric acid or 5 ml concentrated Magnesium, 24 hrs preferred random, if necessary
HCl are acceptable preservatives
Amino Acids, 24 hrs or random Microalbumin Screen, 24 24 hrs or random
Amylase, 24 hrs or random Myoglobin clearance, timed collection. Minimum 4 hours. Refrigerate during
collection. Deliver to lab immediately.
Bence Jones Proteins, random Myoglobin, random
Calcium, 24 hrs preferred; random, if necessary Organic Acids, random
Chloride, 24 hrs or random Osmolality, random
Cortisol, Free, 24 hrs N-Telopeptide, Cross-Linked, 24 hrs
Creatinine, 24 hrs or random Neuroblastoma Screen, random
Creatinine Clearance, 24 hrs Phenolphthalein, 24 hrs or random
Cystine, Quantitative Phosphate, 24 hrs preferred random, if necessary. Refrigerate during collection.
Cytologic Evaluation, random Porphobilinogen, Qualitative, random
D-Xylose, 5 hour only Potassium, 24 hrs or random
Ethanol, random Pregnancy Test, 1st void or random (preferred)
Glucose, Quantitative, 24 hrs or random Protein Electrophoresis, random
Hemocystine, random Protein, Qualitative, random
Hemoglobin, random *3 Protein, Quantitative, 24 hrs or random
Hemosiderin, random Sodium, 24 hrs or random
Immunofixation Electrophoresis, random Urea Nitrogen, 24 hrs or random
Inorganic Phosphorus (Phosphate), random Uric Acid, 24 24 hrs preferred random, if necessary
Lysozyme, random Urobilinogen, random
Urinalysis, Routine Screen, random

Prepared by. DR. Abdulrazzaq Othman Alagbare

 8 Specimen Collection For Culture

Urine Tests Requiring Preservatives or Special Containers

Test name is listed first, with required preservative second. Recommended collection time period is 24 hours, except for pH and
bicarbonate. Sodium, potassium, creatinine, and urea nitrogen can be performed on any routine specimen, with or without
preservative.

17-Hydroxycorticosteroids 1 g boric acid as a preservative added at start of collection

5-Hydroxyindolacetic Acid (5-HIAA) Glass container with 25 mls of 6N HCl
17-Ketosteroids 25 mls 50% HAc; 15 ml 50% HAc for children < 5 years
Aldosterone Refrigerate during collection or 10 grams boric acid or 5 ml concentrated HCl are acceptable preservatives
Aluminum Acid washed plastic container, no preservative
Aminolevulinic Acid, Quantitative Protect from light
Arsenic Plastic container, refrigerate during collection
Bromide Acid washed plastic container, no preservative
Cadmium Acid washed plastic container, no preservative
Carbon Dioxide (CO2 Content) Random container with thin layer of mineral oil
Catecholamines, Fractionated Glass container with 25 mls 6N HCl
Chromium Acid washed plastic container, no preservative
Citric Acid Refrigerated during entire collection period
Copper Acid washed plastic container, no preservative

Prepared by. DR. Abdulrazzaq Othman Alagbare