Urineanalysis 1
Urineanalysis 1
Urineanalysis 1
6. Diagnosis of pregnancy.
COLLECTION OF URINE
First morning, midstream: Preferred for routine urine
examination.
Random, midstream: Routine urine examination.
First morning, midstream, clean catch:
Bacteriological examination.
Postprandial: Estimation of glucose, urobilinogen
24-hour: Quantitative estimation of proteins or
hormones.
Catheterised: Bacteriological examination in infants,
bedridden patients, and in obstruction of urinary tract.
Plastic bag (e.g. colostomy bag) tied around genitals:
Infants, incontinent adults.
CHANGES WHICH OCCUR IN STANDING
URINE@ROOM TEMPERATURE
Increase in pH due to production of ammonia from
urea by urease-producing bacteria.
Formation of crystals due to precipitation of
phosphates and calcium (making the urine turbid)
Loss of ketone bodies, since they are volatile.
Decrease in glucose due to glycolysis and utilization
of glucose by cells and bacteria.
Oxidation of bilirubin to biliverdin causing false-
negative test for bilirubin
Oxidation of urobilinogen to urobilin causing false-
negative test for urobilinogen
Bacterial proliferation
Disintegration of cellular elements, especially in
alkaline and hypotonic urine.
Urine sample must be tested in the laboratory within 2 hours of
collection to get the correct results.
PRESERVATION OF URINE SAMPLE
Refrigeration (4-6°C) is the best general method of
preservation of urine sample up to 8 hours.
Following chemical preservatives can be added to the
24-hour urine sample:
Hydrochloric acid: It is used for preservation of a 24- hour
urine sample for adrenaline, noradrenaline, vanillylmandelic
acid, and steroids.
Toluene: It forms a thin layer over the surface and acts as a
physical barrier for bacteria and air. It is used for
measurement of chemicals.
Boric acid: A general preservative.
Thymol: It inhibits bacteria and fungi.
Formalin: It is an excellent chemical for preservation of
formed elements.
URINE EXAMINATION
PHYSICAL EXAMINATION
CHEMICAL EXAMINATION
MICROSCOPIC EXAMINATION
PHYSICAL EXAMINATION
Volume
Average 24-hr urinary output in adults is 600-2000 ml.
The volume varies according to fluid intake, diet, and climate.
Abnormalities of urinary volume are as follows:
Polyuria means urinary volume > 2000 ml/24 hours.
This is seen in diabetes mellitus, diabetes insipidus,
chronic renal failure, diuretic therapy.
Oliguria means urinary volume < 400 ml/24 hours.
This is seen in febrile states, acute glomerulonephritis, congestive
cardiac failure or dehydration.
Anuria means urinary output < 100 ml/24 hours or complete
cessation of urine output.
This occurs in acute tubular necrosis (e.g. in shock, hemolytic
transfusion reaction), acute glomerulonephritis and complete urinary
tract obstruction.
PHYSICAL EXAMINATION
Color
Normal urine color in a fresh state is pale yellow or amber and is
due to the presence of various pigments collectively called
urochrome.
PHYSICAL EXAMINATION
Appearance
Urinometer
PHYSICAL EXAMINATION
Specific Gravity (SG)
Urinometer
1. Fill a measuring cylinder with 50 ml
of urine.
2. Lower urinometer gently into the
urine and let it float freely.
3. Let urinometer settle; it should not
touch the sides or bottom of the
cylinder.
4. Take the reading of SG on the scale
(lowest point of meniscus) at the
surface of the urine.
5. Take out the urinometer and
immediately note the temperature of
urine with a thermometer.
6. For every 3°C increase / decrease
add / subtract 0.001
PHYSICAL EXAMINATION
Specific Gravity (SG)
Refractometer method:
2. pH indicator paper:
Reagent area (impregnated with bromothymol blue and methyl
red) of indicator paper strip is dipped in urine sample and the
color change is compared with the color guide provided.
Approximate pH is obtained.
PHYSICAL EXAMINATION
Reaction and pH
3. pH meter:
An electrode of pH meter is dipped in urine sample and pH
is read off directly from the digital display.
It is used if exact pH is required.
PHYSICAL EXAMINATION
CHEMICAL EXAMINATION
MICROSCOPIC EXAMINATION
CHEMICAL EXAMINATION
Interpretation-
Appearance of white ring at the junction indicates
presence of protein.
CHEMICAL EXAMINATION
Test for Proteins
Reagent Strip Method
Principle –
The reagent area of the strip
is coated with bromophenol
blue indicator and buffered to
an acid pH which changes
color in the presence of
proteins.
Method –
Bromophenol coated strip is Interpretation -
dipped in urine.
Change in colour of strip
indicates presence of proteins
in urine and is compared with
the colour chart provided for
Reagent strip test is mainly reactive to albumin.
semiquantitative grading.
It is false-negative in the presence of Bence Jones proteins, myoglobin,
CHEMICAL EXAMINATION
Microalbuminuria
Urinary excretion of 30 to 300 mg/24 hours (or 2-20 mg/dl) of
albumin in urine.
Significance –
earliest sign of renal damage in diabetes mellitus (diabetic
nephropathy).
independent risk factor for cardiovascular disease in diabetes
mellitus.
Detection –
Measurement of albumin-creatinine ratio in a random urine sample.
Measurement of albumin in an early morning or random urine
sample.
Measurement of albumin in a 24 hr sample .
Test strips that screen for microalbuminuria are available
commercially.
CHEMICAL EXAMINATION
Quantitative Estimation of Proteins in Urine.
Esbach’s albuminometer method
Fill the albuminometer with urine up
to mark U.
Add Esbach’s reagent (5g picric acid
+ 10g citric acid + 500ml of distilled
water) up to mark R
Stopper the tube, mix it and let it
stand for 24 hours.
Take the reading from the level of
precipitation in the albuminometer
tube and divide it by 10 to get the
percentage of proteins.
CHEMICAL EXAMINATION
Quantitative Estimation of Proteins in Urine.
Turbidimetric method
Take 1 ml of urine and 1 ml standard in two separate
tubes.
Add 4 ml of trichloroacetic acid to each tube.
After 5 minutes take the reading with red filter (680
nm).
CHEMICAL EXAMINATION
Bence Jones Proteinuria
Principle –
When urine is boiled in Benedict’s qualitative solution,
blue alkaline copper sulphate is reduced to red-brown
cuprous oxide if abreducing agent is present.
CHEMICAL EXAMINATION
Test for Glucose
Benedict’s qualitative test:
Methods: Interpretation:
The result is reported in grades
Add 0.5 ml (or 8 drops) of as follows :
urine. Mix well. Nil: no change from blue color
Boil over a flame for 2 minutes. Trace: Green without
Allow to cool at room
precipitate
temperature. 1+ (approx. 0.5 grams/dl):
Green with precipitate
Note the color change, if any. 2+ (approx. 1.0 grams/dl):
Brown precipitate
3+ (approx. 1.5 grams/dl:
Yellow-orange precipitate
4+ (> 2.0 grams/dl): Brick- red
precipitate.
CHEMICAL EXAMINATION
Test for Glucose
Clinitest tablet method (Copper reduction tablet test):
This is a modified form of Benedict’s test in which the
reagents are present in a tablet form.
Sensitivity is 200 mgs/dl of glucose.
CHEMICAL EXAMINATION
Test for Glucose
Reagent strip method
This test is specific for glucose and is therefore preferred over
Benedict’s and Clinitest methods.
It is based on glucose oxidase-peroxidase reaction.
Reagent area of the strips is impregnated with two enzymes - glucose
oxidase and peroxidase and a chromogen.
Method:
Take 5 ml of urine in a test tube and saturate
it with ammonium sulphate.
Add a small crystal of sodium nitroprusside.
Mix well.
Slowly run along the side of the test tube
liquor ammonia to form a layer.
Immediate formation of a purple
permanganate colored ring at the junction of
the two fluids indicates a positive test
CHEMICAL EXAMINATION
Tests For Ketone Bodies
Acetest tablet test
•In acute viral hepatitis, bilirubin appears in urine even before jaundice is
clinically apparent.
2. Gmelin’s test:
Take 3 ml of concentrated nitric acid in a test tube and slowly
placetube
• The equal quantitygently;
is shaken of urine over
play of it.
colors
(yellow, red, violet, blue, & green) indicates
+ve test.
3. Lugol iodine test:
Take 4 ml of Lugol iodine solution (Iodine 1 gm, potassium
iodide 2 gm, and distilled water to make 100 ml) in a test
tube and add 4 drops of urine. Mix by shaking. Development
of green color indicates positive test.
CHEMICAL EXAMINATION
Tests For Detection of Bilirubin in Urine
4. Fouchet’s test: A simple and sensitive test.
i. Add 2.5 ml of 10% of barium chloride to 5 ml of fresh urine in a
test tube and mix well. A precipitate of sulphates appears to which
bilirubin is bound (barium sulphate-bilirubin complex).
ii. Filter to obtain the precipitate on a filter paper.
iii. To the precipitate on the filter paper, add 1 drop of Fouchet’s
reagent (25 g of trichloroacetic acid, 10 ml of 10% ferric chloride,
and distilled water 100 ml).
iv. Immediate development of blue-green color around the drop
indicates presence of bilirubin.
CHEMICAL EXAMINATION
Tests For Detection of Bilirubin in Urine
PHYSICAL EXAMINATION
CHEMICAL EXAMINATION
MICROSCOPIC EXAMINATION
MICROSCOPIC EXAMINATION
Normal urine microscopy contains few epithelial cells,
occasional RBC’s, few crystals.
Urine consists of various microscopic, insoluble, solid
elements in suspension.
These elements are classified as organized or
unorganized.
Organized substances include red blood cells, white
blood cells, epithelial cells, casts, bacteria, and parasites.
Unorganized substances are crystalline and amorphous
material which are suspended in urine and on standing
they settle down and sediment at the bottom of the
containerurinary deposits or urinary sediments.
The cellular elements are best preserved in acid,
hypertonic urine; they deteriorate rapidly in alkaline,
hypotonic solution.
MICROSCOPIC EXAMINATION
Microscopic urinalysis is
done by pouring the urine
sample into a test tube
and centrifuging it for a
few minutes.
The top liquid part (the
supernatant) is discarded.
The solid part left in the
bottom of the test tube
(the urine sediment) is
mixed with the remaining
drop of urine in the test
tube
one drop of this is
analyzed under a
microscope
MICROSCOPIC EXAMINATION
MICROSCOPIC EXAMINATION
Urinary findings in renal diseases
MICROSCOPIC EXAMINATION
Cells in urine
MICROSCOPIC EXAMINATION
Crystals
Crystals are refractile
structures with a definite
geometric shape due to
orderly 3-dimensional
arrangement of its
atoms and molecules.
Amorphous material (or
deposit) has no definite
shape and is commonly
seen in the form of
granular aggregates or
clumps
MICROSCOPIC EXAMINATION
Crystals
MICROSCOPIC EXAMINATION
Crystals
Crystals in acidic urine Crystals in alkaline urine
Uric acid Ammonium magnesium
Waxy casts
represent the final
stage of
degeneration of
cellular casts. They
are more refractile
seen in tubular injury of a more chronic
nature than granular or cellular casts like
severe chronic renal disease and renal
amyloidosis. These casts are also
called renal failure casts.
MICROSCOPIC EXAMINATION
Casts in urine
Fatty Casts
The presence of
red blood cells
within the cast is
always pathologic,
and is strongly
indicative of
glomerular
damage.
They are usually
associated with
nephritic
syndromes.
MICROSCOPIC EXAMINATION
Casts in urine
White blood cell casts