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1.Formation of Urine

2.Normal constituents and composition

3.Collection of urine specimens

4.Preservation of urine

5.Physical examination of urine

6.Chemical Examination of Urine

7. Microscopic Examination Of Urine Deposits

• Urine --- most easily obtained
• Examination of urine
• Information--- about the functioning of
the kidney
• diagnosis ---of urinary tract diseases
• diagnosis -----of certain metabolic and
systemic diseases. 3
• Urine is formed in the kidney.
• The functional unit of kidney is called nephrons,
• where in the ultrafilteration of plasma takes
• followed by absorption of most of the water and
some of the solutes.
• The kidneys through the nephrons ---excrete
many of the waste products of the body 4
Constituent’s grams/litre
• Chloride 9.0
• Phosphorous 2.0
• Sulfur 1.5
• Sodium 4.0
• Potassium 2.0
• Calcium 0.2
• Magnesium 0.2
• Iron 003
• Urea 25
• Uric acid 0.6
• Creatinine 1.5
• Ammonia 0.6
• Sugar not detected by benedicts test trace
• Ketone bodies trace
• Carbonates, bicarbonates & free carbonic acid trace
• Mucin & mucin like substances Diastase 5
• In. Improper collection---- may invalidate the results
• Containers for collection of urine should be wide mouthed, clean
and dry
• First morning sample –concentrated urine ---
chemical constituents
casts and crystals
• Random specimen
chemical screening
microscopic examinations
• 24 hours urine sample
quantitative estimation of proteins, sugars, electrolytes, and
Urine specimen is collected in a clean 2 liters bottle with a
stopper. The first morning sample is not collected.
All the urine passed during the rest of the day and night and next
day I st sample is collected.
Volume is measured and immediately sent to the laboratory. 6
Midstream urine specimen
• Urine should be collected in a clean, dry and,
preferably, sterilized container.
• urethral opening is cleaned with a moist cotton
• first 10 – 25 ml of urine is not collected
• since it contains urethral and prostatic
secretions which may be required if
investigating urethra and prostrate.
• First morning samples ---- these are the
concentrated samples and casts, crystals etc .if
present 7
• Terminal urine sample patient voids the last portion of
urine in an open container.
• Urine specimen collection using a catheter. This
procedure is used for certain bacteriological tests
• Urine specimens from infants
• Urine can be collected into a plastic bag with an
adhesive mouth.
• The bag is fixed around the genitalia and left
in place for 1- 3 hours, depending on the
examination requested.
• Colostomy bags can be used 8

• Urine sample < within 2 hours.

• If delay ----preservation.
• If more than 2 hours
– Keep the sample in the refrigerator without any
– 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. 9

• Volume –
normal -- 1.2-2 L /day
the day is 3-4 times > night.
night is < 400 ml.
polyuria >2000ml / day.
Oliguria <500ml / day.
Anuria is total suppression of urine <100 ml per day.
• Appearance – color & turbidity
• 1. Color - normal ---- amber yellow ( to the presence
of urobilin,uroerythrin,and urochromes) 10
1.Colorless---Very dilute urine
2.Yellow orange(high colored)--Concentrated urine
-----------Excess urobilin
------------Bile pigments
------------Intake of carrots
(Yellow foam ++)
3.Red/ smoky------Hemoglobin/
---- RBC
----MyoglobinBeetroot /
---------- anilinedyes
-------Menstrual contamination
4.Cloudy--------Phosphates & carbonates, Urates & uric acid
Pus cells
5.Milky-------Pyuria,Fat ,chyluria
6. Brown black----

7. orange--------Bile pigments,Drugs – pyridium,rifampicin 11
• 2. Turbidity – normal urine----clear.

• Turbidity in urine may be due to:

 a. amorphous phosphate and carbonates – alkaline or
 neutral urine
 disappears on addition of dilute acetic acid

 b. crystals, cellular exudates, bacteria and fungus

 c. chyle and fat

 d. pus

 e. amorphous urates in acidic urine 12
• b. Odour: Normal-- aromatic odour
On standing--- ammonical
there is decomposition of urea
forming ammonia which gives a strong
ammonical smell..

• Abnormal odour of urine

Fruity---- Ketonuria
Mousy---- Phenylketonuria
Rancid------ Tyrosinaemia 13
• ability to maintain normal hydrogen ion concentration

• Normal kidneys are capable of producing urine the pH of which

varies from 4.5 to slightly higher than 8.0.

• A pH below 7 indicates acid urine and a pH above7 alkaline urine


• Dip the litumus paper strips in the urine, remove and read the color
change immediately.
• Blue litmus turns red - acid
• Red litmus turns blue - alkaline
• Blue and red litmus turns purple - neutral
• nitrazine paper method :The Ph---- nitrazine paper which
are sensitive and specific in the pH range of 4.5 – 8.0 range. 14
• degree of concentration or dilution of the
• specific gravity measures the
concentrating and diluting abilities of the
• The normal --------------1.015 and 1.025
(in a 24 hours specimen).
• urinometer: This is a weighted bulb-
a scale calibrated----- from 1.000 to
1.060 15
Urine is poured into a cylindrical or conical glass so that the vessel is
nearly full.
Froth-- removed with a filter
• The instrument is floated in the urine and care should be taken to
see that it does not touch the slides.

• The depth to which it sinks in urine indicates the specific gravity of


• read on the urinometer scale at the junction of the urine with the

• The reading is taken at eye level, the lowest part of the meniscus
being taken.

• In case the urine is insufficient, it may be diluted with an equal

volume of distilled water and the last two figures of the reading are
then multiplied by 2. 16 17
• Correction for temperature-
• The urinometer is calibrated to read 1.000 in distilled
water at a specific temperature, indicated on each
instrument e.g. 15 C or 20 C.
• There is a change in the specific gravity of 0.001 for
each 3 C above and below this temperature. Therefore
add 0.001 to the reading for each 3 C above the
temperature for which the urinometer is calibrated,
• substract 0.001 for each 3 C, the temperature below the
standard temperature.
• For example - urinometer calibrated for 20 C
• Specific gravity of urine at 32 C is 1.001
• Corrected specific gravity {(32 – 20)/ 3 X 0.001} +
1.001 = 1.005
• Correction is also recommended when glucose or protein
are present. It is recommended that .003 be subtracted
from the urinometer reading for each 1000 mg / dl of
glucose or protein. 18

• This is a non-specific test useful for semiquantitation of marked


• Benedict’s qualitative test:

• This test is not specific for sugars and is affected by most of the
reducing substances.

• Composition of Benedict’s reagent:

Copper sulphate - 17.3 gms
Sodium carbonate (anhydrous) – 100 gms
Crystalline sodium carbonate - 200 Gms
Sodium citrate - 175 gms
Potassium citrate 19
• Dissolve crystalline copper sulphate in 100 ml of
distilled water. Dissolve sodium carbonate and
sodium citrate in 700 cc. of distilled water.
Slowly add the latter to the former solution with
constant stirring. When complete, make up the
volume to 1000ml with distilled water.
• Take 5ml of benedicts reagent
• Boil for 3 – 5 minutes
• add to it 0.5ml (8 drops)of protein free urine.

• Cool and note the color. 20 21
Recording results
The color varies from blue through green –
yellow- orange- brick red.
Negative – no change in color.
Trace - solution appears pale green to slightly
1+ - Definite cloudy green (0.5% sugar)
2+ - Yellow to orange precipitate, supernatant
fluid pale blue (1% sugar)
3+ - Orange to red precipitate, supernatant fluid
pale blue (1.5% sugars)
4+ - Brick red precipitate, supernatant fluid
decolorizes (2 % sugar) 22
• Principle: this test is based on a
double sequential enzyme reaction.
• One enzyme, glucose oxidase, catalyzes
the formation of gluconic acid and
hydrogen peroxide from the oxidation of
• A second enzyme, peroxides catalyzes
the reaction of hydrogen peroxide with
potassium iodide chromogen to oxidize
the chromogen to colors ranging from
green to brown. 23 24
KETONES IN URINE (ketonuria):
• fats--------carbon dioxide and water.
• inadequate carbohydrate in the diet or a defect in
carbohydrate metabolism or absorption,
• fatty acids------------metabolized.
• When the fatty acid utilization is incomplete------- the
intermediary products of fat metabolism appear in the
blood and the urine.
• These products -------- acetone,
• diacetic acid (acetoacetic acid)
betahydroxybutyric acid.
• Diabetes mellitus,
• Other Causes of Ketonuria:
– Fever
– Anorexia
– Gastrointestinal disturbances
– Fasting
– Starvation
– Severe vomiting 25
• Rothera’s Test for Acetone and
Acetoacetic Acid:
• Procedure:
– Take 5ml of urine in a test tube and saturate
it with ammonium sulphate.
– Add 1 crystal of sodium nitroprusside.Run
liquor ammonia carefully at the side of the
tube so as to form a layer on top of the
saturated urine. 26 27
Result: a permanganate calomel red (pink –
purple) ring forms at the junction of the
two layers------positive.
• Negative result shows no ring or a brown
• Ferric Chloride Test for Diacetic Acid –
Gerhardt’s Test
• Harts Test for Beta-Hydroxybutyric Acid
• Colorimetric Reagent Strip Test 28
• 3. Proteins in urine:
Normal <30 mgms / 100 ml (30 – 50
mgms/24 hours)
Tests for Detection of Proteins
• Semiquantitative precipitation tests:
1.The heat and acetic acid method,

2.sulfosalicyclic acid method and

3.the concentrated nitric acid protein

precipitation method precipitation. 29
• Negative – No turbidity or cloudiness.

• Trace - A faint precipitate visible against a

black background, equivalent to about 5 mg / dl
• 1+ - Definite cloud without flocculation
equivalent to 10 – 30 mg / dl.
• 2+ - Heavy and granular cloud without
flocculation equivalent to 40 – 100 mg / dl.
• 3+ - dense cloud with marked flocculation
equivalent to 200 – 500 mg / dl .
• 4+ - cloudiness with precipitation equivalent
to 500 mg / dl or more. 30
• Heat and Acetic Acid Method
Procedure :
• Take a long test tube and fill ¾ the tube with
clear urine.
• Boil the upper portion over a flame .the lower
portion serves as the control.
• If proteins, phosphates or carbonates are
present in the urine a white cloud develops.
• Add 1-3 drops of glacial acetic acid. Any
turbidity due to phosphate precipitation will
clear or if it is due to carbonates they
disappear with effervescence.
• If it persists, it is due to albumin .
• precipitates due to mucin or nucleoproteins will
disappear on addition of 2 drops of nitric acid. 31 32
• Sulphosalicylic acid method
• Procedure
• Take 2 ml of clear urine in a test tube.
• Add an equal volume of 30 %
sulphosalicylic acid.
• Mix thoroughly, allow it to stand for 10
minutes and estimate the amount of
turbidity. 33
• Nitric acid method
• Procedure
• Take 2 – 3 ml of concentrated nitric acid in a
test tube.
• Carefully pour 5ml of clear urine down the
inner side of the inclined test
• tube so that the urine forms a layer over
the nitric acid.
• A ring of white precipitated protein will form
at the interface. Estimates the amount of
precipitate. 34
• Colorimetric reagent strip test
• PRINCIPLE: the colorimetric reagent strip test is based upon the
ability of proteins to alter the color of some acid-base indicators
without altering the pH. When an indicator such as
tetrabromphenol blue is buffered at pH is yellow in solutions
without protein. But in the presence of protein the color will
change to green and then to blue with increasing protein

• PROCEDURE: protein is determined by dipping the strip into well

mixed un centrifuged urine and immediately comparing the
resultant color with the chart provided on the reagent strip bottle.

• RESULTS: the results are reported as negative (yellow color),

trace, 1+ to 4+.trace readings may detect 5 to 20 mg of protein/
dl. Albumin reacts more strongly than the other proteins. Highly
buffered alkaline urine and contamination of the urine specimen
with quaternary ammonium compounds or skin cleansers
containing chlorohexidine may produce false results. 35
Bence Jones Proteinuria:

• Bence Jones protein is soluble at room

and body temperatures.
• It precipitates upon heating between 45 C
and 60 C
• redissolves when the urine is further
heated to the boiling point.

• Reappear on cooling 36
red blood cells

 When hemolysis occurs

 in circulation
 urine
 Normally an occasional red cell may be found
on microscopic examination of the urine
sediment. In women during menstruation, the
urine may get contaminated with menstrual
blood and hence examination of urine should
not be done during that period. 37
• Haematuria: Denotes the presence of red
blood cells in urine. It is seen in various renal
disorders, infectious or neoplastic or trauma
related to any part of urinary tract.

• Hemoglobinuria: is the presence of blood

pigments in the urine without the presence of
red blood cells. It is associated with certain
hemolytic anemia’s that cause hemolytic
anemia, transfusion reactions, malaria, and
paroxysmal nocturnal hemoglobinuria 38
• Tests for Detection of Blood:
• 1. Microscopic Examination:

• 2. Benzidine Test
Heme acts as a catalyst when hydrogen peroxide is
mixed with benzidine.
A: Saturated solution of benzidine in glacial acetic acid
B: Hydrogen peroxide
Mix equal parts of A and B in a test tube and an
equal amount of the mixed reagent.

• A blue color indicates the presence of hemoglobin. 39
Colorimetric reagent strip

Principle: the reagent area is impregnated

with tetramethylbenzidine and buffered
organic peroxide. This forms a green to
dark blue compound when hemoglobin
catalyzes the oxidation reaction of
tetramethylbenzidine with a peroxide. 40
The constituents
1. bilirubin (bile pigments),
2. bile salts,
3. urobilin and urobilinogen.

Bilirubin in appears IN JAUNDICE

Bilirubin in Urine: bilirubin in the urine indicates the presence of

hepatocellular disease or intra or extrahepatic biliary obstruction
in the reticuloendothelial cells

breakdown of hemoglobin.
linked to albumin---------- liver. This albumin-bound form,
which is also known as unconjugated bilirubin( indirect bilirubin)
insoluble in water and does not appear in the urine.
In the liver cells,--conjugated with glucuronic and sulfuric acids
to form water soluble conjugated bilirubin (direct bilirubin).
It is secreted into the bile and then excreted into the intestinal
tract through the bile duct.
This conjugated bilirubin -------------to urobilinogen. 41
• certain liver diseases --unable to
conjugate all the bilirubin-----an increase
in both conjugated and unconjugated
bilirubin --bilirubinuria.

• obstructive biliary tract disease--


• hemolytic anemia’s unconjugated

bilirubin-------- urine free from
bilirubin(Acholuric) 42
Tests for detection of bile salts:
Hay Test
flowers of sulphur --sprinkled on the surface
of the urine,
Results:if bile salts are present they sink to
the bottom.
Otherwise they float on the surface.
This is due to the property of bile salts to
lower surface tension 43
Tests for detection of bile pigments
1. Foam Test:
Shake urine in a test tube. If the foam on
top is yellow, bile pigments are present.
2. Gmelins test:
1. Place ½ inch column of yellow nitric acid
in a test tube.
2. Overlay with equal amounts of urine

A play of colored rings, the most distinct being
green indicates the presence of bile pigments. 44
3. Fouchets Test:
Trichloroacetic acid – 25 gms
Distilled water - 100 ml
10% Ferric chloride solution – 10 ml.
1. Place 10 ml of acidified urine in a test tube.
2. Add 2.5ml of 10 % barium chloride.
3. Mix and filter.
4. Unfold the filter paper and spread it on a dry
5. Add 1 drop of Fouchets reagent to the residual
RESULT: A green or blue color indicates the presence of
bilirubin. 45
Colorimetric Strip Reagent Test
• Principle: this test is base on the coupling of
bilirubin with diazotized 2, 4-
• dichloroaniline in a strong acid
medium to form a brown purple azobilirubin
• Compound. The color ranges
through various shades of tan.
• Procedure: the reagent strip is dipped into
fresh, uncentrifuged urine tapped to
• remove excess urine and after 20
seconds, compared to the color chart
• on the reagent strip bottle
• Result: the results are interpreted as negative,
small (+), moderate (++), and large (+++)
amounts of bilirubin. 46
Tests for detection of
1. Qualitative Ehrlich’s Test
Paradimethylaminobenzaldehyde – 2 gms
20%5 HCL - 100 ml
a. Place 10 ml of urine in a test tube.
b. Add 2.5 ml of barium chloride (to remove
c. Mix well and filter.
d. Add 0.5 ml of Ehrlich’s reagent
e. Allow it stand for 3 – 5 minutes. 47
pink color
observable when viewed from the top of
the test tube. Against a white background
placed beneath the bottom of the test
cherry red color
Abnormally high amounts of This test
must be done with fresh urine or else
urobilinogen is oxidized on exposure to air
urobilin. Excessively cold water should
not be used in diluting
the urine. 48
Qualitative technique:

the urine must be freshly voided

examined without excessive delay in order to

prevent cellular deterioration.

Cellular debris from the urethral meatus and

secretions from the vagina may contaminate the
urine specimen. 49
10-15 ml of urine ----from freshly mixed urine
specimen and centrifuged at a standard speed,
usually 1500 to 2000 rpm for 5 minutes.

This is sufficient to bring to the bottom casts, pus

cells, blood and crystals. For bacteria however a
higher speed of 3,000 rpm is required.

the sediment resuspended in 1 ml.of the same


A drop of resuspended sediment is placed

directly on a microscope slide and covered with
a cover slip.
. 50
low power-
Casts tend to congregate at the edges of the
cover slip.
A minimum of 10 – 15 high power fields should
be scanned for this examination.

• Red blood cells, leucocytes,epithelial cells---

per high power field(/hpf);
• casts --- per low power fields(/lpf).

Other elements such as bacteria, parasites,

crystals and spermatozoa are reported as well 51
Normal sediment contains a limited number
of formed elements. it can be divided into
two classes.

• unorganized sediment

• Organized sediment 52
A. Unorganized sediment - these are the crystals of various
substances present in the urine and they vary with the
pH of the urine .crystals of normal urine is formed as
the specimen cools.
1. Crystals in acidic urine:
a. Uric acid and Urates; –
crystals are seen when the urine is allowed to stand for
sometime and
are not seen in freshly passed urine.
Amorphous urates appear as red granules and are
dissolved by heat and sodium hydroxide but not acetic
Uric acid crystals vary in shape and are yellow brown
in colour and are not dissolved by heat, acetic acid or
HCL but are soluble when heated with sodium
disturbances of uric acid metabolism
fevers where the urine is concentrated. 53
B.Calcium Oxalate:
They are commonly found in diets rich in
tomatoes, spinach etc. they are typically
envelope shaped crystals but occasionally
appear dumb- bell shaped .they are insoluble in
strong Hcl.
c. Cystine Crystals:
highly refractile, hexagonal plates and are
soluble in Hcl but insoluble in acetic acid. They
are seen cystinosis which is an inborn error of
metabolism in which cystine crystals are found in
the urine, reticuloendothelial system and eyes.
slightly yellow, oily looking spheres with radial
and concentric striations .they are not soluble in
Hcl or ether .they are found in liver disorders. 54
• e.Tyrosine:–
fine needles arranged in concentric sheaves, constriction at the
liver disorders.

• f.Sulpha crystals :-
patients taking sulphonamides.

2. Crystals in alkaline urine:

• a. Ammonium Magnesium Phosphates:

( triple phosphate)
coffin lid, feathery or leaf like forms.
In freshly passed urine they indicate stones in the bladder or
Phosphates may occur as amorphous deposits in alkaline urine
and are dissolved in acetic acid.
• b. Dicalcium Phosphates: hey are also seen in slightly acid or
neutral urine. They are colorless prisms arranged in stars and
They are soluble in acetic acid.
c. Calcium carbonate: amorphous granules or colorless spheres 55
and dumb-bells which are soluble in acetic acid with gas formation.
Crystals in acid urine
Amorphous urates-Brick – Red-
Uric Acid -Yellow – Brown--
Polymorphous – Whetstones,Rosettes of
prisms,Rhombohedral prisms, hexagonal
Sodium urateColorless to YellowFan of
slender prisms
Cystine (rare)1. Colorless2.Highly
refractileFlat hexagonal plates with well
– defined edges singly or in clusters 56
Cholestrol (rare) -ColorlessBroken window
panes with notched corners ,Flat plates

Leucine (rare)-1. Yellow or Brown2.Highly

refractile-Spheroids with striations pure form

Tyrosine (rare) Colorless or YellowFine silky

needles in sheaves or rosettes

Bilirubin-Reddish Brown--Cubes, Rhombic

plates, Amorphous needles 57
Acid, Neutral or Slightly
Alkaline Urine

Calcium oxalate--Colorless Octahedral

Dumbbells,Often small

Hippuric acid Colorless- Rhombic plates

Four sided prisms 58
Alkaline Urine
Calcium Carbonate-Colorless

Ammonium biurate-Yellow Opaque

apple,Spheres,Dumbbells,Sheaves of

Calcium Phosphate-Colorless- Prisms,

Plates, Needles - - - - 59
A- represents the residue of normal human urine, as seen under the

B is represented oxalate of urea. An excess of this element indicates


Nitrate of urea is represented in division C. 60
A and B--highly magnified urinary deposits, which indicate different
degrees of impairment of the digestive functions are represented. 61
epithelial cells

In division A is represented pus and mucus, the presence of which

indicates suppuration of the kidneys (Bright's disease). In B pus
globules are alone represented. In the division marked C are shown
blood corpuscles as they are arranged in blood drawn from a vein or
artery. D represents the same separated, as they always are when
present in the urine. In E highly magnified oil globules are represented.
If present in the urine, they indicate disease of the kidneys. In the
division marked F are represented epithelial cells, the presence of
which in large numbers is indicative of disease of the mucous lining of
the urinary organs. 62 63
Uric acid

• In division A are represented the mixed urates as they appear

during idiopathic fevers, as intermittent, remittent, etc. When
appearing as seen in division B, a less violent affection of the same
character is indicated. Division C represents urate of ammonia,
occasionally observed when there is a tendency towards
albuminuria, In division D which is present in the urine of persons
suffering from gout. The crystals shown in division E consist of the
same salt. 64 65 66
varieties of cancer. 67
• Organized Sediment: the components of organized
sediment include casts, red blood cells, white blood cells,
epithelial cells, bacteria, yeast, parasites, spermatozoa
and artifacts.
• a. Casts:
• Casts are formed in the tubules and is composed of
proteinaceous material. They are washed out by the
glomerular secretion into the collecting tubules and the
bladder. They are cylindrical in shape with round or
broken ends with uniform diameter but varying in
length. They require acidic conditions, high salt
concentration, reduced urine flow and protein to be
formed. Practically all casts have a hyaline matrix, which
may or may not contain inclusions such as desquamated
• The casts are named according to the matrix of the
inclusions contained in them e.g. red blood cell casts. 68
1. Hyaline casts:
are colorless,
semi-transparent and
occasionally refractile cylinders and are soluble in acetic
acid. They are seen when there is damage to the
glomerular capillary membrane, permitting leakage of
proteins through the glomerular filtrate. They are seen
in fever, orthostatic proteinuria, and emotional stress or
strenuous exercise.
2. Granular casts: are casts containing large or fine
granules embedded in coagulated protein. They are not
found in normal urine and their presences indicate
pyelonephritis. They are also seen in chronic lead
3. Epithelial casts: are formed of fused desquamated
tubular cells. They are coagulated protein in which are
embedded desquamated epithelial cells from the renal
tubules .they are seen in diseases where there is
damage to the tubular epithelium as in nephrosis,
and heavy metal poisoning. 69
4. Red Blood Cell Casts:
are casts with red blood cells embedded
in the coagulated protein in the tubule.
Their presences indicate acute
inflammation or vascular disorder in the
glomerulus causing hematuria. They are
seen in pathological conditions such as
acute glomerulonephritis, renal infarction
and collagen vascular disorder.
5. White Blood Cell Casts (Pus cell)
6. Fatty Casts
7. Waxy Casts 70
• a. Red blood cells: Normally 1-2 red blood cell
are found per high power field
• .they appear pale , light refractive, biconcave
discs when viewed under high power
magnification .they have no nuclei. Red blood
cells in fresh, unstained sediment appear pale in
color; in urine that is not fresh, they are pale or
colorless shadow cells .in concentrated urine,
they may be small and crenated; and in dilute
urine, they are large and swollen and sometimes
rupture to produce ghost cells.
• b. White cells
• c. Epithelial Cells: Normally a few epithelial
cells occur in the urine .A marked increase
• is these cells in the urine is seen destruction
of the tissues in
urinary tract. 71
• Quantitative Evaluation of the urine
sediment – Addis count
• The Addis count is a quantitative
measurement of the excretion of red cells,
leucocytes and casts in the urine during a
12 hour period.
• e. Bacteria:
Bacteruria is considered significant when
there is the presence of 100,000 or more
bacteria per ml of urine specimen. 72
• Microscopic Examination:
sediment - >20 or more bacteria per high
power field may indicate a urinary tract infection
Reagent strips:
This test depends upon the conversion of nitrate
to nitrite by the action of gram negative bacteria
in urine. At the acid pH of the reagent area,
nitrite in the urine reacts with p-arsanilic acid to
form diazonium compound. This compound in
turn couples with 1, 2, 3, 4-tetra
hydrobenzoquinolin-3-ol to produce pink color.
• Procedure: The strip is dipped in the urine
specimen for 5 seconds. 73
uniform pink color --positive result

the presence of 100,000 or more organisms per ml,

A negative result should never be interpreted as indicating absence of


There are several reasons for this.

– First morning urine or urine that has remained in the bladder for
several hours is more likely to yield a positive nitrite test result
in the presence of significant bacteruria than a random urine
sample that may have been in the bladder for short time. in the
latter type of specimen there may have been insufficient
time(less than 4 hours) for conversion of nitrate to nitrite to
have occurred.
– When dietary nitrate is absent, even if organisms containing
reductase are present and bladder incubation is ample.
– Ascorbic acid concentrations of more than 25 mg / dl or greater
may cause false negative results with specimens containing
small amounts of nitrate. 74 75