MTAMBANI LABORATORY AND GENERAL CLINIC HEALTH SERVICES

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Document No:02

STANDARD OPERATING PROCEDURE FOR URINALYSIS

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1. Purpose

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 This procedure provides instructions for examination of Urine.
2. Scope
All ……… laboratory technical staff working at Parasitology section
3. Responsibility
 It is the responsibility of the section head to ensure implementation of this SOP
4. Principle
The appearance of urine colour, turbidity and form is noted before performing chemical
and microscopical examination. Commercially produced reagent strips are the most
convenient method for detecting chemical substances in urine. Strips are impregnated
with chemicals and indicators which undergo a colour change if the target substance is
present in urine. An estimate of the quantity of the substance is determined from the
depth of colour change produced (semi –quantitative reaction).Strips are available that
detect a few or multiple chemical substances
5. Materials

Reagent Supplies Equipment

 Iodine  Microscopically slide  Light Microscope
 Normal saline  Lens paper  Centrifuge machine
 Eosin  Urine container
 Pipettes
 Cotton gauze,
6. Sample and container type
Containers for the collection of urine should be wide-mouthed, clean and dry. If the urine
specimen has to be transported for any length of time it should contain an appropriate
preservative to prevent bacterial overgrowth or hatching of viable ova.

7. Environmental and safety controls

 All specimens must be regarded as potentially infections.
 Never leave urine specimens exposed to the air or sun light in containers without
lids.
 Never accept urine specimens mixed with stool (e.g. in a bedpan).
 Never examine urine specimens without first putting on gloves.

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  Always examine urine specimens within 1–2 hours after collection.
8. Calibration:
Use calibrated centrifuge.
9. Quality Control

 Run known negative and positive specimens daily or whenever a new strip bottle
is first opened.
10. Procedure

10.1 Collection of urine specimens

 Containers for the collection of urine should be wide-mouthed, clean and dry. If
the urine specimen has to be transported for any length of time it should contain
an appropriate preservative to prevent bacterial overgrowth or hatching of viable
ova
10.2 Types of urine specimen
 Early morning urine specimen: Early morning urine provides the most
concentrated sample.
 Random urine specimen: A random urine sample, taken at any time of the day,
will enable the laboratory to screen for substances which are indicators of
kidney infection.
 24-Hour urine specimen: The 24-hour urine specimen is collected in a clear 2-
litre bottle with a stopper. On the first morning the patient gets up and urinates;
this urine is not collected. All the urine passed during the rest of the day and
night is collected in the bottle. The next morning the patient gets up and collects
the first urine of the morning in the bottle. The bottle should then be taken
immediately to the laboratory. Measure the volume of urine with a measuring
cylinder and record it.
 Midstream urine specimen: While passing urine, the patient places an open
container in the stream of urine and collects about 20 ml of urine. The container
should be covered immediately.
 Terminal urine specimen: The patient urinates the last portion of urine into an
open container.

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  Urine specimens collected using a catheter: Collection of urine using a
catheter must be carried out by a qualified physician or nurse. The procedure is
used for certain bacteriological tests, mainly in women. Usually, however, a
specimen collected in the normal way following thorough cleansing is
acceptable for this purpose
10.3 Urine collection:
 Whenever possible, the first urine passed by the patient at the beginning of
the day should be sent for examination. This specimen is the most
concentrated and therefore the most suitable for culture, microscopy and
biochemical analysis
 Give the patient a sterile, dry, wide-necked, leak proof container and request
10-20 ml of urine specimen.
 Explain to the patient the need to collect the urine with as little
contamination as possible i.e. a clean-catch specimen
 Female patients: Instruct them to cleanse the area around the urethral
opening with clean water, dry the area, and collect the urine with the labia
held apart.
 Male patients: Instruct them to wash their hands before collecting a
specimen (middle of the urine flow).
 Label the container with the date, the 3 initials and ID of the patient, and
time of specimen collection. As soon as possible, deliver the specimen with a
request form to the laboratory
 When immediate delivery to the laboratory is not possible, refrigerate the
urine at 4-6°C When a delay in delivery of more than 2 hours is anticipated,
add boric acid preservative to the urine
10.4 Urine chemical test
 The dipsticks are placed into the urine and immediately removed. They are
then compared with a comparison chart after an appropriate time that is also
specified on the chart. The colour changes observed on the dipstick will give
a semi-quantitative estimation of the amount of substance present. This can
be reported as negative, +, ++,+++, ++++ or as an approximate value of the
concentration of the substance tested

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  Put 10 mls of well mixed urine sample into the centrifuge tube
 Centrifuge at medium speed 2000rpm for 2 minute
 Pour the supernatant fluid (by completely inverting the tube) into a second
container not the original one.
 This can be used for biochemical tests to avoid contaminating the original
urine which may need to be cultured (depending on the findings of the
microscopically examination
 Dip all the test pads of the strip into the urine and immediately remove the
strip .if reading the strip visually start timing
 Drag the edge of the strip against the container rim to remove excess urine
and blot the edge on a paper towel or tissue. If reading visually
 Compare each test pad to the corresponding row of colour blocks on the
bottle label
 Read each pad at the time shown on the label starting with the shortest time
 Hold the strip close to the colour blocks and match carefully
 Reads the pads in good light
 If using an analyzer; Place the test strip on the analyzer according to the
analyzer operating manual. The analyzer will automatically reads each test
pad at specified time
10.5 Microscopic Examination of Urine.
 Urine is examined microscopically as a wet preparation to detect significant
pyuria i.e. WBCs in excess of 10 cells/ml (106/l) of urine
 Red cells, Casts, Yeast cells, Trichomonas vaginalis, motile trophozoites,
Schistosoma haematobium eggs, Bacteria (providing the urine is freshly
collected)
10.6 Preparation and examination of a wet preparation
 Aseptically transfer about 10 ml of well mixed urine to a labeled conical tube.
 Centrifuge at 1000rpm for 5 minutes.
 Pour the supernatant fluid (by completely inverting the tube) into a second
container not the original one.

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  This can be used for biochemical tests to avoid contaminating the original urine
which may need to be cultured (depending on the findings of the microscopically
examination
 Remix the sediment by tapping the bottom of the tube.
 Transfer one drop of the well mixed sediment to a slide and cover with cover
glass
 Note: Do not discard the remaining sediment because this may be needed to
prepare a Gram smear if WBCs and or bacteria are seen in the wet preparation.
 Examine the preparation microscopically using the 10x and 40x objective with
the condenser iris closed sufficiently to give good contrast.
10.7 Deterioration of urine
The following changes occur when unpreserved urine is left at room
temperature:
 Any bacteria in the urine will multiply so that the bacterial count will be
unreliable.
 When the organisms are urease-producing, the ammonia released will increase
the pH of the specimen which will result in the destruction of cells and casts.
Bacteria will also break down any glucose which may be present
 When white cells, red cells, and casts are present, these will begin to lyze
especially in a concentrated specimen.
 The concentration of the bilirubin present may be oxidized to biliverdin which
will not be detected. Likewise, urobilinogen will not be detected because it will
be oxidized to urobilin.

Appearance Possible Cause

Cloudy. Urine usually has unpleasant Bacterial Urinary Infection

smell and contains WBCs

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 Red and cloudy due to red cells Urinary Schistosomiasis Bacterial
Infection
Brown and cloudy due to Haemoglobin Black water fever Other
conditions that cause intravascular Hemolysis
Yellow-Brown, or green-brown due to Acute Viral Hepatitis Obstructive
Bilirubin
Jaundice
Yellow-orange due to urobilin i.e. Hemolysis
Oxidized uribilinogen Hepatocellular jaundice
Milky-white due to chyle Bancroftian filariasis

11. Results reporting

 Report the macroscopic appearance,
 Ova/egg seen and all other abnormal findings.
 Count the number of wbcs/hpf (x40 objective)
12. References
 Parasitology Atlas and Health Laboratory Safety
 Monica Cheesbrough, Medical Laboratory Manual for Tropical Countries
Volume I
 Manual of basic techniques for a health laboratories Second edition-WHO

AMENDMENT SHEET

Revision date Amendment Version Reviewed by Next review date

Staff acknowledgement documentation log

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I do hereby acknowledge that I have read and understood the content of this document.
N° Staff Name Staff signature Date

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