MT- Micro 1 (Lab) Routine Urine Culture

Catheterizing the Urinary Bladder for Collection Aim of the test

 Another method is the catheterized urine specimen in which a
lubricated catheter (thin rubber tube) is inserted through the
urethra (tube-like structure in which urine is expelled from the
bladder) into the bladder. This avoids contamination from the
urethra or external genitalia.
 An etiological diagnosis of bacterial urinary tract infection by semi
quantitative cultivation of the urine with identification and
susceptibility test of the isolated bacteria(s).
Types of specimen

 Urine (Midstream urine), suprapubic aspiration, catheterized

urine
 Note: First morning specimens yield highest bacterial
counts from overnight incubation in the bladder, and
are the best specimens.

Criteria of specimen rejection

Preservation and Storage
Changes that affect the
chemical or
microscopic properties
of urine is kept at room
temperature for more
than 1 hour
 Un-refrigerated specimen older than 2 hours may be subject to
overgrowth and may not yield valid results, unlabeled specimen,
Refrigeration- most mislabeled specimen, and specimen is expired transport
common method for container, 24 hours urine specimens.
storing and preserving
urine Pathogens and commensals
-It prevents bacterial
growth for 24 hours
-After 24 hours use
chemical preservation

Specimen Collection and Transport Guidelines

 All urine collection and/or transport containers should be clean

and free of particles or interfering substances
 The collection and/or transport container should have a secure lid
and be leak-resistant. Leak-resistant containers reduce specimen
loss and healthcare worker exposure to the specimen while also
protecting the specimen form containers

Urine Specimen Handling Guidelines

 Labels include the patient name and identification on labels.

Make sure that the information on the container label and the
requisition match. If the collection container is used for transport,
the label should be placed on the container and not on the lid,
since the lid can be mistakenly placed on a different container. Pre specimen processing
Ensure that the labels used on the containers are adherent under
Patient preparing
refrigerated conditions.
 Collection Date and Time includes collection time and date on the
 Instruct the procedures for the patient Specimen collection
specimen label. This will confirm that the collection was done
 Collection of midstream urine for bacterial investigation:
correctly. For timed specimens, verify start and stop times of
 Patient not needing assistance:
collection. Document the time at which the specimen was
 Give the patient a suitable container.
received in the laboratory for verification of proper handling and
 Instruct the patient before the collection, preferably with
transport after collection
illustration.
 Tell the patient not to touch the inside or rim of the container
Prompt transportation needed

Who will collect the specimen?

 Transport to lab ASAP
 Urine should be within 2 hours or be refrigerated for up to 24
 Midstream urine is collected by the patient
hours
 If disabled, nursing staff will assistant in collection
 Microorganisms grow very rapidly in urine at room temperature
 For catheterized specimen, nursing staff will collect the specimen
 This could give a false positive culture result
 Suprapubic aspiration is performed by the physician
Quantity of specimens
 To fill line in transport tube (-20ml)
Time relapse before processing the sample
 The maximum time allowed for processing a urine sample is 2
hours from the time of collection
Storage
 At room temperature unless delay is inevitable; it must be
refrigerated or mixed with preservative like boric acid
Specimen Processing
URINE
WET MOUNT GRAM STAIN
BLOOD AGAR MACCONKEY AGAR NUTRIENT AGAR
URINE
Initial report
WET MOUNT
 The use of dipstick designed to detect the presence of urine
nitrite and to indirectly estimate the number of segmented
neutrophiles through the detection of leukocyte esterase activity.
Rationale for the nitrate test I tat most urinary tract infections are
caused by nitrate reducing members of the family
Enterobacteriaceae.
 Leukocyte Esterase (LE) is produced by segmented neutrophiles;
this test when perfomed alone correlated with ten or more white
blood cells per high power field in the urine equates with a
sensitivity in the range 88% and specificity 94%
Screening Test URINE
GRAM STAIN
 As many as 60% to 80% of all urine specimens received for culture
by the acute care medical center laboratory may contain no
etiological agents of infection. Procedure developed to identify
quickly those urine specimens that will be negative on culture,
thus to circumvent excessive use of media, technologist time, and
the overnight incubation period. The gram stain is the easiest,
least expensive, and probably the most sensitive and reliable
screening method for identifying urine specimen that contain
greater that 105 CFU/ml
 A drop of well-mixed in urine is allowed to air dry
 The smear Is stained and examined under of
immersion (1000X)
 Presence of at least one organism per oil immersion
field (examining 20 fields) correlates with significant
bacteriuria (> 105 CFU/ml)
URINE
BLOOD AGAR MACCONKEY AGAR NUTRIENT AGAR
Culturing Procedure
 Mix the urine sample to re-suspend microorganism present.
 Dip a 1 μl or 10 μl calibrated loop in vertical position in the urine
and remove the loop and use the collected fluid to inoculate
Nutrient, Blood and MacConkey agars respectively.
Colony Counting
 CFU/ml In original sample = # colonies counted/ (Dilution)
(Volume plated in mL)
 A plate count of 100,000 CFU/ml of pure culture should be
considered positive and isolated organism should be identified
and sensitivity test will be performed.
 A plate count between 10,000-100,000 CFU/ml is considered
suspected
 A plate count between less than 10,000 CFU/ml is considered
negative.
Post specimen processing
Interfering factors
 Patient on antibiotic therapy
 Improper sample collection
Result reporting
 Report wet amount as an initial report
 Report the isolated pathogen and its sensitivity pattern as a final
report
Turn around time
 Wet mount results should be available 1 hour after specimen
report
 Isolation of a possible pathogen can be expected after 2-3 days
 Negative culture will be reported out 1-2 days after the receipt of
the specimen
Additional Information
 A single culture is about 80% accurate in the female, two
containing the same organism with count of 10’5 or more
represents 95% chance of true bacteriuria., three such specimens
mean virtual certainty of true bacteriuria.
 Single clean voided specimen from an adult made may be
considered diagnostic with proper preparation and care in
specimen collection
 UTI is significantly higher in woman who use diaphragm
spermicide contraception, perhaps secondary to increased vaginal
pH and a higher frequency of vaginal colonization with E.coli
 Most laboratories limit the number of organisms which will be
identified when recovered from urine to two.
 If the patient is receiving antimicrobial therapy at the time the
specimen is collected, any level of bacteriuria may be significant
 Failure to recover aerobic organisms from patients with pyuria or
positive Gram’s stains of urinary sediment may indicate the
presence of mycobacteria or anaerobes.