LESSON 2: INTRODUCTION TO URINALYSIS

URINALYSIS

• Analyzing urine was actually the beginning of laboratory medicine

• It is the oldest laboratory procedure performed
• According to CLSI, testing of urine with procedures commonly performed in an expeditious,
reliable, accurate, safe and cost-effective manner.

What are the reasons why we perform urinalysis?

• It aids in the diagnosis of disease

- It is useful in ascertaining evidence of disease or disturbed functions of the kidneys and also
helpful in drug screening
• It screens asymptomatic populations for undetected disorders
- Ex. Determining inborn errors of metabolism
• It monitors the progress of disease and effectiveness of therapy.
- Common practice when monitoring treatment of diabetes which involves many daily urine
test for urinary glucose excretion

• The color, odor and amount of urine can already indicate whether something is wrong
- Ex: Dark urine – could be a sign that they are dehydrated, or their kidneys are not working
properly
• A complete urinalysis is composed of multiple tests including physical, chemical, and
microscopic analysis or examination.

HISTORY OF URINALYSIS

1. Laboratory medicine began 6,000 years ago with the analysis of human urine, which was called
uroscopy until the 17th century and today is termed urinalysis.
• So today, Physicians use urine to diagnose selective conditions but from Ancient times until the
Victorian era, urine was used as the primary diagnostic tool.
• So Physician spoke of urine as a divine fluid or a window to the body.
2. Babylonia and Egyptian physicians began the art of uroscopy.
• ‘uroscopia’ means scientific examination of urine
• It derives from the Greek word ‘auron’ meaning urine and ‘skopeo’ meaning to examine or
inspect.
• Edwin Smith Surgical Papyrus
- Found in Egyptian hieroglyphics (cave)
- The early physicians examined a bladder-shaped flask of urine
- observe a diagnostic information of urine such as urine color, turbidity, odor, volume,
viscosity and even sweetness.
- 2 Ways To Determine the Sweetness of Urine:
 a) testing test (tinikman nila)
b) attraction of ants in certain specimen
3. Sumerian and Babylonian physicians of 400 BC recorded their assessment of urine on clay tablets.
- urine characteristics were altered with different diseases
4. Sanskrit medical works from 100 BC describe 20 different types of urine.
5. Hindu cultures
- urine tasted sweet and that black ants were attracted to sweet urine, a characteristic of the
disease now known as diabetes mellitus.
6. In the fourth century BC, Hippocrates hypothesized that urine was a filtrate of the humors, which
came from the blood and was filtered through the kidneys.
- 4 humors: blood, phlegm, yellow bile and black bile
- Hippocrates
o describes bubbles that lay on the surface of fresh urine as an indication of long-term
kidney disease.
o bubbles on the surface of urine are in fact often owing to proteinuria (presence of
protein in urine)
o wrote a book in uroscopy
- Galen
- redefined Hippocrates’ ideas, theorizing that urine represented is not a filtrate of the
four humors and overall condition, but rather, a filtrate of the blood.
7. During the middle ages, physicians concentrated with “uroscopy” by means of examining urine and
also part of their training.
8. By 1140 CE, color charts has been developed that describes the significance of 20 different colors.
- Protospatharius
o invented the first documented laboratory technique
o heat would precipitate proteins causing proteinuria to manifest through cloudiness
- Frederik Dekkers
o In 1694, he made a laboratory findings of albuminuria by boiling urine, which
remains a useful ay diagnostic indicator today.
- Paracelsus
o used vinegar to bring out cloudiness
o acid will precipitate or cook proteins
9. By the 17th century, the uses of uroscopy had spiraled far beyond the edge of reason. Physicians
and leches started telling fortunes and predicting the future with urine, a practice known as
‘uromancy’. Witch hunte mixed urine with nails to distinguish witches from on witches. The abuses
of urine finally caused a backlash.
- Thomas Bryant
o led a medical rebellion against all uses of uroscopy over the centuries
o In 1627, he published the Pisse Prophets, a book that devastated uroscopy.
- Thomas Addis
o developed methods for quantitating the microscopic sediment ay
- Richard Bright
o introduced the concept of urinalysis as part of doctor’s routine patient examination
in 1827
Importance of Urinalysis
- Useful in ascertaining evidence of disease or disturbed functions of the kidneys and the
pathological lesions of the uterus, bladder, ureters, for males lesions of prostate and
seminal vesicles.
- Its chemical changes can indicate early disease.
- Common practice of monitoring treatment of diabetes which involves multiple daily urine
test for urinary glucose excretion.
- Frequent study of urine chloride excretion has been proposed as a means of monitoring low
salt diet.
- Determining inborn errors of metabolism.
Urine

- A complex aqueous solution of organic and inorganic constituents resulting from the active
metabolism of the body or directly from foods take in. Is an ultra-filtrate of plasma that carries
most of the bodies waste products and excess water out of the body.
- Two unique characteristics of a urine specimen:
1. It is readily available and easily collected specimen.
2. Urine contains information, which can be obtained by inexpensive laboratory tests,
about many of the body’s major metabolic functions.

Composition of Urine

1. Water - 90%
2. Solutes – 5% (organic and inorganic)

COMPOSITION AND CONCENTRATION OF URINE DEPENDS ON:

1. Dietary intake
Ø For example, Pag concentrated ang urine mo very yellow siya minsan nag tuturn siya
ng orange meaning konti lang yung intake mo ng water kaya mas maraming solutes
ang nag aappear sa urine mo.

2. Physical activity
Ø For example, Yung person had a vigorous exercise so need ng muscles niya ng mas
maraming energy, so more phosphocreatine ang macacatabolize, as a result more
waste product ang magegenerate into the form of creatinine. So tataas ang
creatinine sa urine ng person ng nagvivigorous exercise.

3. Body metabolism
Ø For example, if there is an increase nucleic acid metabolism meaning tataas ang uric
acid mo, lalabas na ito sa urine mo.

4. Endocrine function
Ø For example, the patient from to is suffering diabetes mellitus, ang main problem
wala siyang insulin to carry the glucose into the cell, so the tendency of that is tataas
 yung blood glucose level niya. Since hindi maka-pasok ang glucose sa loob ng cell
yung excess glucose ay lalabas sa urine niya.

5. Body position (orthostatic)

Ø For example, Orthostatic proteinuria eto yung pag increase ng protein excretion sa
urine during upward position bet decrease during horizontal position or naka higa
ka.
Ø Tumataas din ang protein enzyme mo

ORGANIC COMPONENTS 14. Hormones

1. Urea — 25-35g/
2. Creatinine — 1.5g/
INORGANIC COMPONENTS
3. Uric Acid — 0.4-10g/L
4. Hippuric Acid — 0.7g/L 1. Chloride — 10g/L
5. Undetermined N2 — 0.6g/L 2. Sodium — 5g/L
6. Ketones 3. Potassium — 3.3g/L ;
7. Carbohydrates 4. Calcium — 0.3g/L
8. Bicarbonates 5. Magnesium — 0.1/L
9. Sugar 6. Sulfate -—2.5g/L
10. Pigments - 2.9g/L 7. Phosphate — 2.5g/L
11. Mucin 8. Ammonium — 0.7g/L
12. Fatty acids 9. Phosphorous — 2¢/L
13. Enzyme 10. Total Sulfur — 1.5g/L

Two Types of Solutes

Organic Components Inorganic Components

Urea – major organic Chloride – major inorganic

Creatinine NaCl

Uric acid Potassium

Hippuric acid Sulfate, phosphate, ammonium, calcium and

magnesium
Other substances such as CHO, pigments, fatty
acids, mucins, enzymes and hormones *highest to lowest concentration

Organic Components

Urea

• Major organic component present in urine

• 60-90% nitrogenous material
 • Product of protein and amino acid metabolism
• Approximately 6-18 grams per 24 hours appear in the urine

Creatinine

• Product of creatinine metabolism by muscles

• Approximately 0.3-0.8 grams per 24 hours appear in the urine

Uric Acid

• Product of nucleic acid breakdown in food and cells

• Approximately 0.08-0.2 grams per 24 hours appear in the urine

Inorganic Components

Chloride

• Approximately 100-250 milli equivalent per 24 hours appear in urine

Sodium chloride (NaCl)

• Principle salt
• Approximately 100-200 milli equivalent per 24 hours appear in urine

Potassium (K)

• Combined with chloride and other salts

• Approximately 50-70 milli equivalent per 24 hours appear in urine

Phosphate

• Combined to sodium to buffer the blood

Ammonium

• Regulates blood and tissue fluid acidity

Calcium

• Combines with chloride, sulphate and phosphate

3 MAJOR ORGANIC COMPONENTS (highest to 3 MAJOR INORGANIC COMPONENTS (highest to

lowest concentration) lowest concentration) CHaNaK

1. Urea 1. Chloride
2. Creatinine 2. Sodium
3. Uric acid 3. Potassium

To verify if that a specimen is urine:

ü Urea and Creatinine content is being tested because these substances are present in much higher
concentrations approximately 50% of plasma in urine than in other body fluids.
 ü Values of about 600 mg per deciliter Urea Nitrogen and 50 mg per deciliter Creatinine indicates
urine.
ü Urine also contains hormones, vitamins, medications and formed elements such as cells, cast,
bacteria, crystals and mucus which may increase indicates presence of disease.

URINE COLLECTION
1. Specimen must be collected in clean, dry, leak-proof containers
2. Properly applied screw-top lid are less likely to leak than snap-on lids
3. Containers for routine analysis
a. Wide mouth and a wide, flat bottom
• To prevent over turning
b. Made of clear material
• To allow determination of color and clarity
c. Recommended for capacity is 50 mL
• 12ml in the urine is needed for microscopic analysis
d. For microbiologic urine studies, individually packaged sterile containers with secure closures
should be used
e. Sterile containers are also suggested if more than 2 hours’ elapse between specimen
collection and analysis
4. All specimen must be properly labelled with the patient’s name, identification number, date and
time of collection and additional information such as the patient’s age, location and the
physician’s name.
- Take note:
o The label should be attach to the container and not to the lid to prevent
misidentification.
5. Specimen sample delivered to the laboratory should be accompanied with a properly labelled
laboratory request form
- Take note:
o Information on the form must match to the information on the specimen label. The
time the specimen receive in the laboratory should be recorded on the form.

SPECIMEN REJECTION

- Improperly labelled and collected specimens should be rejected by the laboratory and request
for new specimen.

Causes of specimen rejection in urinalysis:

1. Specimens in unlabeled containers
2. Non-matching specimen label and requisition forms
3. Specimen contaminated with feces or toilet paper
4. Containers with contaminated exteriors
5. Specimen of insufficient quantity (QNS)
6. Specimens that have been improperly transported
SPECIMEN HANDLING

- Changes in urine composition takes place is not only in vivo but also in vitro thus requiring
correct handling after specimen collection
- Take note:
- Never discard specimen before checking with a supervisor. Put on the refrigerator
when still processing the problem or error that encountered in the laboratory.

SPECIMEN INTEGRITY

- After collection, specimen must be sent to the laboratory and be tested within 2 hrs.
- Specimen that cannot be delivered and tested within 2hrs, should be refrigerated or have an
appropriate chemical preservative added.
- If not properly preserved, the following changes can be occur:

CHANGES IN UNPRESERVED URINE

Analyte Change Cause

Color Modified/Darkened Oxidation or reduction of

metabolites

Clarity Decreased Bacterial growth and

precipitation of amorphous
material

Odor Increased Bacterial multiplication causing

breakdown of urea to ammonia

PH Increased Breakdown of urea to ammonia

by urease-producing
bacteria/loss of CO2

Glucose Decreased Glycolysis and bacterial use

Ketones Decreased Volatilization and bacterial

metabolism

Bilirubin Decreased Exposure to light/

 Photo oxidation to biliverdin

Urobilinogen Decreased Oxidation to urobilin

Nitrite Increased Multiplication of nitrate-

reducing bacteria

Red and White cells and casts Decreased Disintegration in dilute alkaline
urine

Bacteria Increased Multiplication

Trichomonas Decreased Loss of motility, death

NOTE:

PABAON KO – analytes that increases during unpreserved urines

• pH • Nitrite
• Bacteria • Color
• Odor (THE REST AY DECRESEASES NA)

URINE PRESERVATION

- Urine specimen must be examined immediately for accurate result of tests and for proper
evaluation. Examination may be delayed if specimen can be ideally preserved.
- Refrigeration - most routinely used method of preservation. (Physical preservative not Chemical
preservative)
- Refrigerator Temperature is 2 to 8 Degrees Celsius
- Refrigeration will decrease bacterial growth and metabolism, but it will cost precipitation of
amorphous urease and amorphous phosphate.
• Amorphous Urease – pink precipitate
• Amorphous Phosphate – white precipitate
- Refrigeration it increases specific gravity when measure by urinometer
- If the urine is cultured, it should be refrigerated during transit and kept refrigerated until
cultured up to 24 hrs.
Take note:
• Specimens must be return to the room temperature before chemical testing by reagent
strips because the enzymes reaction on the strips perform best at room temperature
and also this will correct specific gravity and may dissolve some of amorphous urease.
• Strips ay hindi priniprigenate and sensitive sila sa light.
- Room Temperature – 20 to 24 Degrees Celsius

Purposes Of Urine Preservation

• To suppress bacterial growth by:

a. preventing conversion of urea to ammonia
b. preventing degradation of glucose by bacteria or yeast
c. interference of bacterial proteins
 • Kapag present kasi ang bacterial proteins natin, nagkakaroon tayo false positive
protein test
d. Preventing instability of urinary solutes
e. Preventing degeneration of organized sediments such as pus casts, etc.

Ideal Preservative

1. Should be bactericidal
Bactericidal - destroys bacteria
2. Inhibits urease
- Urease is an enzyme that catalyzes the hydrolysis of urea. Its breakdown into carbon dioxide
and ammonia which causes elevation of the urine ph. It means tumataas ang ating ph ng urine,
yung ang iniiwas ng ating preservative.
So that, our ideal preservative ay inhibit ng urease.
3. Preserved formed elements in the sediments
- It is important for diagnosis for microscopic examination of urine
4. Should not interfere with chemical tests

NOTE: Choose preservative that best suits the needs of the required analysis.

Urine Preservatives

Preservatives Advantages Disadvantages Additional Information

Refrigeration Does not interfere with Raises specific gravity Prevents bacterial
chemical tests by hydrometer growth 24 h3

Precipitates amorphous
phosphates and urates

Thymol Preserves glucose and Interferes with acid

sediments well precipitation tests for
protein

Boric Acid Preserves protein and May precipitate crystals Keeps ph at about 6.0
formed elements well when used in large
amounts

Does not interfere with

Is bacteriostatic (not
routine analyses other
bactericidal) at 18 g/L;
than ph
can use for culture
transport

Interferes with drug

and hormone analyses.
 Formalin Excellent sediment Acts as a reducing Rinse specimen
preservative agent, interfering with container with formalin
(Formaldehyde)
chemical tests for to preserve cells and
glucose, blood, casts.
leukocyte, esterase and
copper reduction

Toluene Does not interfere with Floats on surface of

routine tests specimens and clings to
pipettes and testing
materials

Sodium Fluoride Prevent Glycolysis Inhibit reagent strip May use sodium
tests for glucose, blood, benzoate instead of
and leukocytes fluoride for reagent
Is a good preservative strip testing.
for drug analyses

Take note:

For culture and routine analysis – Boric Acid and Thymol are best preservative for urine specimen

Preservatives Advantages Disadvantages Additional Information

Phenol Does not interfere with Causes an odor change Use 1 drop per ounce
routine tests of specimen

Commercial Convenient when May contain one or Check tablet

preservative tablets refrigeration not more of the composition to
possible preservatives including determine possible
sodium fluoride effects on desired tests
Have controlled
concentration to
minimize interference

Urine Collection Kits Contains collection

(Becton Dickson, cup, C&S (culture and
Rutherford, NJ) sensitivity)
preservative tube or
UA(urinalysis) tube

Gray C&S (culture and Sample stable at room Decreases pH; do not Preservative is boric
sensitivity) tube temperature (RT) for use if urine is below acid and may not be
48 hr; preserves minimum fill line used for UA
bacteria

Yellow plain UA tube Use an automated Must refrigerate within Round or conical
instrument 2 hours bottom
 Cherry red/yellow top Stable for 72 hours at Bilirubin and Preservative is sodium
tube RT; instrument- urobilinogen may be propionate; conical
compatible decreased if specimen bottom
is exposed to light and
left at RT

Saccomanno Fixative Preserves cellular Used for cytology

elements studies

Special Preservatives:
1. 10 mL 40% Formalin – Addis count
2. 10 mL conc. HC1 – epinephrine, cathecholamines, vanilymandelic acid (VMA)
3. 10 mL Glacial HAC, pH 2.0 – aldosterone
4. H2SO4 - preserves calcium and other inorganic constituents
5. NaF or Benzoic acid – ideal for glucose analysis; prevents glycolysis

METHODS OF URINE COLLECTION

1. Bottle Method
• A method that uses any receptacles/container to collect the specimen provided that it is
dry, clean and sterile.
2. Gauze-pad Method
• A gauze pad is used to collect the urine and then centrifuged in a centrifuge tube
containing a golf tee.
• For infants
3. Midstream Clean Catch Method
• Collection of urine specimen for examination at the middle part of a single continued
normal urination
• Gitnang ihi na hindi nag iistop because there is a tendency especially for girls na
masama yung epithelial cells sa private area
4. Catheterized Method
• Rubber tubed which has been cleaned and sterilized is inserted through the urethral
orifice to the urethral canal, then finally to the bladder to collect a presumably pure
urine specimen.
• Not recommended anymore because it is painful
5. Suprapubic Aspiration Method
• Direct puncturing of the suprapubic region for collection of urine from the urinary
bladder
• Because the bladder is sterile under normal conditions, suprapubic aspiration provides a
sample for bacterial culture that is completely free of extraneous contamination. The
specimen can also be used for cytologic examination.

Pictures of Catheterized Method and Suprapubic Aspiration

Method
TYPES OF URINE SPECIMEN

1. Single/Random Specimen
• Any urine sample collected any time of the day
• Most commonly received specimen in the lab because it is ease of collection and
convenience
• The actual time of voiding should be recorded on the container.
• Useful for routine screening tests to detect obvious abnormalities.
• Affected by dietary intake or physical activity before collection.
• Advantage: Allows detection of pathologic postprandial (collection after a meal)
concentration of solutes like sugar, protein, carbohydrates
2. First Morning Specimen
• Voided upon rising or waking
• Most ideal screening specimen
• Best for urine determination and the contents of sediments
• prevents false-negative pregnancy test
• Most preferred sample because the nitrate sample is less variable in dilution
• Concentrated urine sample: Detection of chemicals and formed elements that may not
be present in a dilute random specimen.
• Elevates orthostatic proteinuria or postural proteinuria which is characterized by
elevated protein excretion while in the upright position
• The patient should be instructed to collect the specimen immediately on arising and to
deliver it to the laboratory within 2 hours or keep it refrigerated.
3. 24-Hour or Timed Specimen
• Used to produce accurate quantitative results
Note: Used for quantifying analytes and creatinine clearance
• Required for analytes that exhibits diurnal variations and are affected by changes
brought by daily activities such as exercise, meals and body metabolism.
• Note: Solutes that exhibit diurnal variations:
• Catecholamines
• 17 hydroxy steroids
• Electrolytes which are lowest in the early morning and highest in the afternoon
(Sodium and Potassium)
• Procedure: Have the patient completely empty the urinary bladder. The urine is
discarded. Collect all urine samples voided during the next 24 hours including the last
samples voided exactly at the conclusion of the 24-hour period
• Note: The patient must begin and end collection period with an empty bladder.

SAMPLE 24-HOUR (TIMED) SPECIMEN COLLECTION PROCEDURE

Provide the patient with the proper collection contained and preservative

DAY 1: 7 AM – patient voids and discards specimen; collects all urine for the next 24 hours

DAY 2: 7 AM – patient voids and adds this urine to previously collected urine
On arrival at laboratory, the entire 24-hour specimen is thoroughly mixed, and the volume is
measured and recorded

Note:

ü Measure the volume accurately

ü If failed to discard the first voided urine it will cause falsely elevate result
ü If failure to include urine produce at the end of the collection period it will cause falsely
decreased results
ü All the specimens should be refrigerated or kept on ice during the collection period or
may require additional chemical preservative
ü If a specimen is collected into two containers the contents of the containers should be
combined and thoroughly mixed before aliquoting.
4. 12-Hour Urine Sample
• Urine is collected within 12 hours (8am to 8pm)
• Use Addis Count (Developed by Thomas Addis)
• The Addis count quantitates formed elements in a 12-hour overnight urine collection
• Note: If any of these cell’s changes was noted, it indicates a disease process:
§ Casts > 5,000
§ RBC > 500,000
§ WBC > 2,000,000
• Choice of Preservative for Addis Count: 10% Formalin

5. Early Afternoon Specimens

• For urobilinogen determination

• Because urobilinogen excretion is enhanced in alkaline urine the specimen of choice for
quantifying or monitoring is a 2-hour collection following the mid-day meal or lunch is it
between 2 to 4 for pm for early afternoon specimens
• This collection correlates with the typical alkaline time which happens 2 to 4 pm

6. 2-Hour Post Prandial

• Collected shortly before consuming routine meal and collecting again 2 hours after
eating
• For monitoring insulin therapy in diabetic patients

7. Catharized Specimen

• Collected under sterile conditions by passing a catheter through the urethra into the
bladder
• Used to measure function of individual kidney
• Note: Specimens are collected separately by passing catheters through the ureters on
each kidneys not recommended because it is painful
• Most commonly used for bacterial culture
• Note: If a routine urinalysis is also requested the culture should be performed first to
prevent contamination of the specimen. Culture before routine urinalysis
8. Glucose Tolerance Test
• collected to respond with the blood samples drawn during an OGTT.
- For glucose monitoring, may oral glucose tolerance test, nagdradraw ka ng sample
also nagcocollect ka rin for urine sample
• tested for glucose and ketones reported along with the blood test results as an aid to
interpret the patients ability to metabolize a measured amount of glucose.
- May include fasting half hour, 1hour ,2-hour,3-hour specimens. Collection is still
institution’s option.
9. Fasting Specimen (Second Morning)
• second voided specimen after a period of fasting
• the fasting specimen does not contain any metabolites from food ingested before the
beginning of the fasting period
• recommended for glucose monitoring

10. Suprapubic Aspiration Specimen

• It is collected by internal introduction of a needle through the abdomen into the bladder
• It provides a sample for bacterial culture that is completely free of extraneous
contamination because the bladder is sterile under normal condition
• used for cytologic examination
11. Midstream Clean-Catch Specimen
• alternative to catheterized specimen
• safer, less traumatic method of obtaining urine ay for bacterial culture and routine
urinalysis and less contaminated by epithelial cells and bacteria
Note: Lesser yung sakit o trauma sa patient
• more representative of the actual urine than the routinely voided urine
• Routine urinalysis and Culture analysis are requested on a catheterized specimen, the
culture first to prevent contamination of the specimen
Note: Pag midstreamà Gitna (center) ang daloy ng ihi na at di nagsstop to prevent
contamination of asample by epithelial cells of girls
a) Clean-Catch Specimen Collection: Female Cleansing Procedure:
1. Wash hands.
2. Remove the lid from the container without touching the inside of the container or
lid.
§ to avoid contaminating the sterile container
3. Separate the skin folds (labia).
4. Cleanse from front to back on either side of the urinary opening with an antiseptic
towelette, using a clean one for each side.
5. Hold the skin folds apart and begin to void into the toilet.
6. Bring the urine container into the stream of urine and collect an adequate amount
of urine. Do not touch the inside of the container or allow the container to touch
the genital area.
7. Finish voiding into the toilet.
8. Cover the specimen with the lid. Touch only the outside of the lid and container.
 9. Label the container with the name and time of collection and place in the specified
area or follow institutional policy.
§ Label the containers not on the lid
b) Clean-Catch Specimen Collection: Male Cleansing Procedure:
1. Wash hands.
2. Remove the lid from the sterile container without touching the inside of the container
or lid.
3. Cleanse the tip of the penis with antiseptic towelette and let dry. Retract the foreskin if
uncircumcised.
4. Void into the toilet. Hold back foreskin if necessary.
5. Bring the sterile urine container into the stream of urine and collect an adequate
amount of urine. Do not touch the inside of the container or allow the container to
touch the genital area.
- do not touch inside of the container and clean your private area
6. Finish voiding into the toilet.
7. Cover the specimen with the lid. Touch only the outside of the lid and container.
8. Label the container with the name and time of collection and place in the specified area
or follow institutional policy.
c) For Midstream Clean-Catch Specimen:
1. Appropriate cleansing material
- strong bacterial agents such as hexachlorophene or povidone-iodine SHOULD NOT BE
USED.
- Mild towelettes are RECOMMENDED.
2. Sterile containers
3. Instructions for cleaning and voiding

12. Prostatitis Specimen (Three-Glass Collection)

• for prostatic infection determination
• Procedure:
o 1st specimen- first portion of urine
o 2nd specimen- midstream
o 3rd specimen- massaged prostate
§ For third specimen, prostate is massaged first so that the prostate fluid
will be pass with the remaining urine.
• Quantitative cultures are performed on all specimens
- Take note that first and third specimen are for microscopic examination while the
second specimen is use as control for bladder and kidney infection. If the second
specimen is positive the result from the third specimen are invalid because infected
urine has contaminated the specimen.
• (+) Prostatic Infection: 3rd specimen have a WBC/HPO and bacterial count 10 times that
of the first specimen
- Macrophages containing lipids also be present if you have prostatic infection.
• Variations of the procedure
- Stamey - Mears Four Glass Localization method.
 a) Bacterial cultures of the initial voided urine or VB1 - tested for urethral infection
or inflammation
b) Midstream urine or VB2 - tested for urinary bladder infection
c) Express Prostatic Secretion or EPS - cultured and examined for WBC
-Take note that greater than 10-20 WBC per high power field is considered
abnormal
d) Post-prostatic massage urine specimen or VB3 - prostatic massage urine
specimen
• Variations of the procedure:
• Pre and Post massage Test (PPMT)
o clean-catch midstream urine is collected
o second urine is collected after the prostate is massaged
o (+) result : If bacteriuria in the post massage specimen is 10 times the pre-
massage count

13. Pediatric Specimens

• Used a soft, clear plastic bag with hypoallergenic skin adhesive for urine collection
• sterile specimens may be obtained by catherization or by suprapubic aspiration
• Take note specimens for culture may also be obtained using a clean-catch cleansing
procedure and sterile collection bags
• check then applied bags approximately every 15 minutes until the needed amount of
sample has been collected (one of the hardest in pediatric specimen)

DRUG SPECIMEN COLLECTION

• Chain of custody/evidence - Process that provides documentation of proper sample

identification from the time of collection to the receipt of laboratory results.
• In COC, all personnel handling the specimen must be noted.
• Forms of tampering specimen includes substitution, adulteration or dilution
• URINE SPECIMEN COLLECTION FOR DRUG SPECIMEN MAYBE WITNESS OR UNWITNESS

DRUG URINE SPECIMEN

• Volume: 30-45 ml
• Temperature: 32.5 - 37.7 Celsius
o -must be taken within 4 minutes to confirm that the specimen has not been adulterated
• Urine temperature outside the recommended range may indicate specimen contamination.
Color is inspected to identify any signs of contaminants. The sample should be water-free urine.
Bluing agent is added to toilet water to prevent specimen adulteration. If the specimen is
suspected that had been diluted or adulterated, a new sample must be collected, and a
supervisor is notified.

URINE DRUG SPECIMEN COLLECTION PROCEDURE

1. The collector washes hands and wears gloves.

2. The collector adds bluing agent (dye) to the toilet water reservoir to prevent an adulterated
specimen.

3. The collector eliminates any source of water other than toilet by taping the toilet lid and faucet
handles.

4. The donor provides photo identification or positive identification from employer representative.

5. The collector completes step 1 of the chain-of-custody (COC) form and has the donor sign the form.

6. The donor leaves his or her coat, briefcase, and/or purse outside the collection area to avoid the
possibility of concealed substances contaminating the urine.

7. The donor washes his or her hands and receives a specimen cup.

8. The collector remains in the restroom but outside the stall, listening for unauthorized water use,
unless a witnessed collection is requested.

9. The donor hands specimen cup to the collector. Transfer is documented.

10. The collector checks the urine for abnormal color and for the required amount (30 to 45 mL).

11. The collector checks that the temperature strip on the specimen cup reads 32.5°C to 37.7°C. The
collector PROCEDURE 2-4 records the in-range temperature on the COC form (COC step 2). If the
specimen temperature is out of range or the specimen is suspected of having been diluted or
adulterated, a new specimen must be collected and a supervisor notified.

12. The specimen must remain in the sight of the donor and collector at all times.

13. With the donor watching, the collector peels off the specimen identification strips from the COC
form (COC step 3) and puts them on the capped bottle, covering

both sides of the cap.

14. The donor initials the specimen bottle seals.

15. The date and time are written on the seals.

16. The donor completes step 4 on the COC form.

17. The collector completes step 5 on the COC form.

18. Each time the specimen is handled, transferred, or placed in storage, every individual must
be identified and the date and purpose of the change recorded.

19. The collector follows laboratory-specific instructions for packaging the specimen bottles and
laboratory copies of the COC form.

20. The collector distributes the COC copies to appropriate person

TYPES OFURINE SPECIMEN

Types of Specimen Purpose

Random Routine screening
First morning Routine screening
Pregnancy tests
Orthostatic protein
Fasting (second morning) Diabetic screening/monitoring
2-hour postprandial Diabetic monitoring
Glucose tolerance test Optional with blood samples in glucose
tolerance test
24-huor timed) Quantitative chemical test
Catheterized Bacterial culture
Midstream clean-catch Routine screening
Bacterial culture
Suprapubic aspiration Bladder urine for bacterial culture
Cytology
Three-glass collection Prostatic infection