Chapter Two
Collection and Preservation of Urine Specimen
•Introduction
Proper specimen collection is essential for test results
to be valid
Good quality assurance requires that specimens be
collected with care to avoid contamination and to help
prevent erroneous results
Proper handling and transport of specimens after
collection ensures that the laboratory receives a
specimen of acceptable quality for testing
Types of Urine Specimen and Methods of obtaining
•Methods of obtaining Urine Specimen
The patient should be instructed to void directly into a clean, dry
container, or a clean dry bed pan so that the specimen can be
transferred to an appropriate container
Specimens from infants and young children can be collected in a
disposable collection apparatus
If a urine specimen is likely to be contaminated with vaginal
discharge or menstrual blood, this period has to be avoided and
the patient must be informed to bring a clean voided specimen
All specimens should be immediately covered and taken to the
laboratory
Types of Urine Specimen
Random Specimen: Is a specimen obtained at any time during
examination
Is most convenient type of specimen
Is most common type of specimen
Obtained any time during the day
Is good for
Chemical screen
Microscopic Examination
First-Morning Specimen: Is a specimen obtained during the first
urination of the day
Is the preferred type of specimen for most routine urine testing
Is most concentrated
• Is bladder incubated assuring detection of chemicals and formed
elements that may not be present in a dilute random specimen
Is best for testing:
Nitrite
Protein
Microscopic examination
Second Voided Specimen: Is a type of urine specimen where the first
morning specimen is discarded and the second specimen is collected
Is good mainly for
Glucose monitoring
2 hours Postprandial : Is a specimen obtained 2 hours after meal
Good for glucose
Catheterized Specimen
This specimen is collected under sterile conditions by passing a hollow
tube (catheter) through the urethra into the bladder.
The most commonly requested test on a catheterized specimen is a bacterial
culture.
Mid -Stream Clean-catch Urine
Specimen:
• Is an alternative to the catheterized specimen
• Provides a safer, less traumatic method for obtaining urine for bacterial
culture and routine urinalysis.
• Is a specimen obtained from the middle part of the first urine and after
the patient first clean their external genitalia with an antiseptic wipe
• Procedure
• The genital area should be cleaned with soap and water and rinsed well.
This is to keep off bacteria on the skin from contaminating the urine
specimen
• The patient should urinate a small amount and this is discarded
• The urine that comes next, the midstream specimen, should be collected
into a sterile container of 30 to 50ml
– Kits are also available in the market which contains towelettes and a sterile
disposable urine container for the collection of clean-catch urine specimen
24-Hour (or Timed) Specimen
When the concentration of the substance to be measured changes
with diurnal variations and with daily activities such as exercise,
meals, and body metabolism, 24-hour collection is required.
Necessary for quantitative chemical tests like protein, creatinine,
urea etc.
All specimens should be refrigerated or kept on ice during the
collection period and may also require addition of a chemical
preservative.
On its arrival in the laboratory, a 24-hour specimen must be
thoroughly mixed and the volume accurately measured and
recorded.
•
Procedure for Collection of 24-Hour Urine
Specimen
•
1. Inform the patient to completely empty his/her bladder and discard
his/her urine exactly at the beginning of the 24 hour time collection
( lets say at 6:00 a.m)
2. Collect all urine voided during the following 24 hours, including that
voided exactly at the end of the 24 hour period in a container ( at 6:00
a.m) of the following ( second ) day
3. All the urine collected must be preserved
4. The container should be labeled with:
The test order
The patient name
Time of collection
The preservative added
Sources of Error in the Collection of Urine
1.Bacteriological or chemically contaminated specimen
2.Wrong type / amount of preservatives
3.Partial loss specimen or inclusion of two morning
specimens in the 24 hr collection
4.Inadequate mixing of specimen before examination
5.Careless measuring of the 24hr urine volume
•
Handling and Preserving Urine Specimen
•Urine Specimen Containers
Random and first morning specimens may be collected in
clean, lidded, disposable containers large enough to hold at
least 50ml
They should be made of a clear material to allow for
determination of color and clarity.
For clean-catch specimens, urine containers must be sterile
Large opaque containers capable of holding four or more
liters are provided to patients for collecting 24 hr urine
specimens. These containers may contain preservatives
•Labeling Urine Specimens
Urine specimens must be labeled clearly with
the patients name, date and time of collection
Labels should be on the container, not just on
the lid
Preservation of Urine Specimens
Once urine is collected, it should be transported to the laboratory
immediately for analysis (within2 hours).
Over time, the physical and chemical composition of urine change and
formed elements deteriorate.
If it cannot be analyzed immediately, sample should be preserved
appropriately. In short term, refrigeration will slow deterioration of
urine, but there is no good substitute for immediate examination.
Long standing of urine at room temperature can cause;
Rapid growth of bacteria which breaks down urea to ammonia leading to an
increase in pH and precipitation of calcium and phosphate
Lysis of RBCs, WBCs and casts
Oxidation of urobilinogen to urobillin
Destruction of glucose by bacteria
PRESERVATION OF URINE
• Urine sample < within 2 hours.
• If delay ----preservation.
• If more than 2 hours
• –Keep the sample in the refrigerator without any
preservative.
• –Toluene – add a few drops till it forms a thin layer on
the surface of urine.
• –Conc. HCL – 1 ml of conc.HCL for 125-150mlof urine.
• –Formaldehyde -1 drop for 15 ml of urine. Cells and
casts are well preserved.
Method of Preservation of Urine Specimens
Urine can be preserved by physical or chemical methods
a. Physical Means
Refrigeration
Freezing
b. Chemical Method
Thymol
Toluene
Formaldehyde
Hydrochloric Acid ( HCL)
Chloroform
Boric Acid
Chlorhexidine
Sodium carbonate
N.B: No preservative of any kind should be added to urine for bacteriological culture
Types of Examination in Routine Urinalysis
Physical Examination of Urine
Volume
Color
Odor
Appearance
pH
Specific Gravity
Chemical Examination of Urine
Glucose
Protein
Ketone
Bilirubin
Urobillunogen
Nitrite
Leukocyte Esterase
blood
Microscopic Examination of Urine
RBC
WBC
Epithelial Cells
Casts
Bacteria
Yeasts
Parasites
Crystals
Artifacts
•
Categories of Urine Tests
Screening (Qualitative) Tests
Screening tests tell only whether a substance is present or
absent and the results are reported as positive or negative
They are done on random specimen
Semi-Quantitative Tests
Qualitative tests give rough estimate of the amount
of substance present. They are also called semi-
quantitative tests. The results of qualitative tests
can be graded as negative, trace, +1, +2, +3 or +4
Quantitative Tests
Quantitative tests determine accurately the amount of the
substances to be tested. However, since they are time
consuming they are not included in routine urinalysis
Most common quantitative tests performed in urinalysis
laboratory are for protein
The results of a quantitative test usually reported in milligrams
per deciliter, gram per deciliter, per liter
For quantitative test a complete 24 hour urine specimen is
needed. An appropriate preservative should be added to the
container or the specimen should be stored in refrigerator
QUIZ
• Write down at least three types of urine specimen
• Why do preferable first morning specimen than
others
• What is the advantage of random specimen
• Write down at least five types of chemical
examination
• Write down the three types of Examination in
Routine Urinalysis
