Urinalysis Practicum

Clinical Pathology Dept of Medical Fac

Brawijaya Univ/Saiful Anwar
General Hospital Malang
 Sampling Considerations

Wide opening container, capped, clean, dry, not

always sterile except for bacterial culture.
Avoid detergent/desinfectant residues
Minimize contamination
Labelled (patient’s name, registered no, age, sex,
date and time collection)
Must be send to laboratory as soon as possible &
in 1 hour must be examined
If delayed, samples are stored in 4oC
Avoid sunlight exposure
Washing hand Open the Throw first Cap the
container and voided urine, container
place the cap then fill the without
faces on top container as touching inside
needed of the cap
The types of urine specimen
 Wanita
• Pasien dianjurkan mencuci tangan dengan air
bersih.
• Duduk, paha dibuka, dengan jari bibir
kemaluan dibuka pisah, sehingga lubang
keluar urin tampak.
• Cuci dengan air bersih.
• Urin dikeluarkan.
• Pancaran permulaan dibuang, berikutnya
porsi tengah ditampung.
 Pria
• Pasien dianjurkan mencuci tangan dengan
air bersih.
• Pada mereka yang tidak bersunat, kulup
kemaluan ditarik ke belakang, sehingga
lubang keluar urin tampak, cuci dengan air
bersih.
• Urin dikeluarkan.
• Pancaran permulaan dibuang, berikutnya
porsi tengah ditampung.
 Per kateter
• Kateter dipasang aseptis.
• Pancaran tengah ditampung dalam
botol bersih dan steril.
• Pada kateter tinggal menggunakan
jarum injeksi tabung semprit,
ditusuk diisap ditampung dalam
penampung bersih steril.
 Per pungsi suprapubik
• Pasien dianjurkan memenuhkan
kandung seninya.
• Kulit 3 jari supra pubik didesinfeksi.
• Menggunakan jarum semprit, ditusuk,
diisap urinnya.
• Jarum ditutup, urin dalam semprit
dikirim.
Urine preservatives

Refrigerator
Boric acid
Thymol
40% formaldehide
Sodium florida
Toluen
Changes in unpreserved urine
 Sample Processing
Sentrifuge 1500-2000
rpm, 5 min

10-12 ml
Urine

For chemical For microscopic (0.5

examination cc)
 Sentrifuge 1500-2000
rpm, 5 min

10-12 ml
urine, mix:
for chemical
dipstick

For microscopic (0.5 cc)

 Urine Protein (Boiling method)
3-5 drops
3-6% acetic
acid
- : Clear
+1 : slightly cloudy
+2 : Cloudy, seed
+3 : Cloudy,
fragmented
2 ml
Urine +4 : Cloudy,
agglutinate
Test control
 Interpretation

Negative +1 +2 +3 +4
Glucose urine (reduction test)

• Principle:
Glucose will reduce cupri
(Cu3+) to cupro (Cu2+)
 Benedict method
8 drops
Urine Negative Positive 3+

5 ml
Reagent

Boil
 Interpretation

Negative + 1 +2 +3 +4
Negative (-) : Blue or greeny blue, cloudy
Positive 1+ : yellowish green and cloudy
Positive 2+ : yellow -green, cloudy
Positive 3+ : orange, cloudy
Positive 4+ : brownish red
 Procedure

1. Test urine as soon as possible after

receipt (use fresh urine)
2. Remove only enough strips for
immediate use; recap tightly.
3. Test a well-mixed, unspun urine
sample.
4. Do not touch the test area with
fingers.
5. Dip reagent strip into urine briefly –
no longer than 1 second.
6. Drain excess urine off – run edge of strip
along rim of tube
7. Touch the edge of the strip to an
absorbent paper
8. Do not lay reagent strip directly on
workbench surface
9. Follow exact timing recommendations for
each chemical test. Usually 60 sec, no
more than 2 min
10. Hold reagent strip close to the color chart
and read under good lighting.
Urine Multistix – reading dipstick results
manually; colors are matched to those
on the bottle label; timing is critical for
each pad.
 Specific gravity

1. Polyelectrolyte: polymethylvinyl ether/maleic acid

2. Indicator: bromthymol blue

• Polyelectrolyte  sensitive to ions in urine

• If electrolytes level is increased, SG is
increased too
 pH

Manufacturers use a double-

indicator system of methyl red
and bromthymol blue.

Methyl red H → Bromthymol blue

H
(Red-Orange → Yellow) (Green →
Blue)
 Protein

The protein area of the strip

contains either
tetrabromphenol blue or 3′, 3′
′, 5′, 5′′-tetrachlorophenol-3,
4, 5, 6-
tetrabromosulfonphthalein
and an acid buffer to maintain
the pH at a constant level.
Readings are reported in terms of negative,
trace, 1+, 2+, 3+, and 4+; or the semiquantitative
values of 30, 100, 300, or 2000 mg/dL
corresponding to each color change.

Negative

Positive
 Source of error/interference
False positive False negative
Highly buffered alkaline Proteins other than
urine albumin
Pigmented specimens, Microalbuminuria
phenazopyridine
Quaternary ammonium
compounds (detergents)
Antiseptics, chlorhexidine
Loss of buffer from
prolonged exposure of the
reagent strip to the
specimen
High specific gravity
 Glucose

glucose oxidase
1. Glucose + O2 (air) → gluconic acid H2O2

peroxidase
2. H2O2 + chromogen → oxidized colored chromogen + H2O
 Interference
• False-positive:
Contamination by oxidizing agents
and detergents
• False-negative:
High levels of ascorbic Acid,
High levels of ketones
High specific gravity
Low temperatures
Improperly preserved specimens
 Keton

• Results are reported qualitatively as negative,

trace, small (1+), moderate (2+), or large (3+),
or semiquantitatively as negative, trace (5
mg/dL), small (15 mg/dL), moderate (40
mg/dL), or large (80 to 160 mg/dL).
alkaline
acetoacetate + sodium nitroprusside + (glycine) →
(and acetone) purple color
 Blood
hemoglobin
H2O2 + chromogen → oxidized chromogen + H2O
peroxidase green-blue color

• False-positive: Bacterial peroxidases,

Menstrual contamination
• False-negative: High specific gravity/crenated
cells, Formalin, Captopril, High concentrations
of nitrite, Ascorbic acid 25 mg/dL, Unmixed
specimens
 Bilirubin

acid
bilirubin glucuronide + diazonium salt → azodye

• False-positive: Highly pigmented urines,

phenazopyridine, Indican (intestinal
disorders), Metabolites of Iodine
• False-negative: Specimen exposure to light,
Ascorbic acid 25 mg/dL, High concentrations
of nitrite
 Urobilinogen

acid
urobilinogen + p-dimethylaminobenzaldehyde → red color
(Ehrlich’s (Ehrlich reagent)
reactive
substances)
• False-positive: Porphobilinogen, Indican, p-
aminosalicylic acid, Sulfonamides, Methyldopa,
Procaine, Chlorpromazine, Highly-pigmented urine
• False-negative: Old specimens, Preservation in
formalin
 Leukocyte esterase
leukocyte esterases
indoxylcarbonic acid ester → indoxyl + acid indoxyl
acid
+ diazonium salt → purple azodye
• False-positive: Strong oxidizing agents,
Formalin, Highly pigmented urine,
nitrofurantoin
• False-negative: High concentrations of protein,
glucose, oxalic acid, ascorbic acid, gentamicin,
cephalosporins, tetracyclines, inaccurate
timing
 Nitrite

acid
para-arsanilic acid or sulfanilamide NO2 → diazonium salt
(nitrite)
acid
diazonium salt tetrahydrobenzoquinolin → pink azodye

P-arsanilic
acid

positive
 Result Reporting
• Procedures:
1. Protein (boiling method)
2. Glucose (Benedict)
3. Dipstick
• Chemical manually:
– Protein :
– Glucose :

• Chemical stick:
– Glucose: ...
– Protein : ...
– Keton :… etc.