2. Introduction
Urine is formed in the kidneys, is a product
of ultrafiltration of plasma by the renal
glomeruli.
3. Collection of urine
Early morning sample-qualitative
Random sample- routine
24hrs sample- quantitative
Midstream sample-UTI
Post prandial sample-D.M
Catheterised- in infants, bedridden
patients
Suprapubic needle aspiration
4. 24 hour urine sample
1. For quantitative estimation of proteins
2. For estimation of vanillyl mandelic acid,
5-hydroxyindole acetic acid,
metanephrines.
3. For detection of hormones in urine
4. For detection of microalbuminuria
6. Preservation of urine sample
HCl- for 24 hr urinary sample preservation
for adrenaline, noradr, VMA, steroids.
Toluene- as a physical barrier
Boric acid- general preservative
Thymol- inhibits bacteria, fungi
Formalin- for preservation of formed
elements
7. For routine analysis, preservatives should
be avoided as they interfere with reagent
strip technique and chemical test for
proteins.
Sample should be examined within 1-2 hrs
of voiding.
If delay is expected, sample can be kept in
refrigeration for max 8 hrs.
10. Urinary volume
Normal = 600-2000ml with night urine not in
excess of 400 ml.
Polyuria- >2000ml/24 hrs
Oliguria-<500ml/24 hrs
Anuria-complete cessation of urine(<200ml)
Nocturia-excretion of urine by a adult of >500ml
with a specific gravity of <1.018 at night
(characteristic of chronic glomerulonephritis)
13. Color & appearance
Normal= clear & pale yellow due to the
presence of various pigments called
urochrome.
1. Colourless- dilution, diabetes mellitus,
diabetes insipidus, diuretics
2. Milky-purulent genitourinary tract
infection, chyluria
3. Orange- urobilinogen
4. Red-beetroot ingestion,haematuria,
hemoglobinuria
5. Brown/ black- alkaptunuria, melanin
14. Urinary pH/ reaction
Reaction reflects ability of kidney to
maintain normal hydrogen ion
concentration in plasma & ECF
Normal= 4.6-8
Tested by- 1.litmus paper
2. pH paper
3. Reagent strip method
15. Acidic urine
Ketosis-diabetes, starvation, fever
Systemic acidosis
UTI by E.coli
Acidification therapy
High protein diet
16. Alkaline urine
Strict vegetarian
Systemic alkalosis
UTI by pseudomonas or Proteus
Alkalization therapy
CRF
17. Odour
Normal= aromatic due to the volatile fatty
acids
Ammonical – bacterial action(E. coli)
Fruity- ketonuria, starvation
Musty- Phenylketonuria
Fishy- UTI with Proteus
Rancid- Tyrosinemia
18. Specific gravity
Depends on the concentration of various
solutes in the urine.
Normal range- 1.003 to 1.035
Measured by-urinometer
- refractometer
- reagent strip method
- falling drop method
19. Urinometer
This method is based on the principle of
buoyancy.
Take 2/3 of urinometer container with urine
Allow the urinometer to float into the urine
Read the graduation at the lowest level of
urinary meniscus
Correction of temperature & albumin is a
must.
Urinometer is calibrated at 15or 200
c
So for every 3o
c increase/decrease add/subtract
0.001
22. Low specific
gravity(hyposthenuria)
All causes of polyuria except gycosuria
DI, pyelonephritis, glomerulonephritis.
Fixed specific gravity (isosthenuria)=1.010
Seen in chronic renal disease when kidney
has lost the ability to concentrate or dilute
23. Osmolality
Normal adult with normal fluid intake wil
produce urine of 500-850 mOsm/kg water.
The normal kidney is able to produce
urine osmolality in the range of 800-1400
mOsm/kg water in dehydration and
minimal osmolality of 40-80 mOsm/kg
water during diuresis.
25. Tests for proteins
Test – HEAT & ACETIC ACID TEST
Principle-proteins are denatured & coagulated
on heating to give white cloud precipitate.
Method-take 2/3 of test tube with urine, heat only
the upper part keeping lower part as control.
Presence of phosphates, carbonates, proteins
gives a white cloud formation. Add 1-2 drops of
10% acetic acid, if the cloud persists it indicates
it is protein(acetic acid dissolves the
carbonates/phosphates)
26.
27.
28.
29. Other Tests for Protien
Nitric acid test
Sulphosalicylic acid test
Test with Esbach’s reagent
Protienreagent strip
Biuret method
30. Protein % of Total Daily Maximum
Albumin 40% 60 mg
Tamm-Horsfall 40% 60 mg
Immunoglobulins 12% 24 mg
Secretory IgA 3% 6 mg
Other 5% 10 mg
TOTAL 100% 150 mg
Proteins in “Normal” UrineProteins in “Normal” Urine
31. Causes of proteinuria
Glomerular proteinuria: due to increased
permeability of glomerular capillary wall.
Selective( only albumin n transferrin bands seen)
Nonselective: pattern same as serum
e.g. nephrotic syndrome
Tubular proteinuria: in acute n chronic
pyelonephritis, heavy metal poisoning, TB
kidney etc.
32. Overflow proteinuria: Bence jones
proteins(plasma cell dyscrasia),
hemglobin( intravascular hemlysis),
myoglobin(skeletal muscle trauma)
Hemodynamic proteinuria: seen in high
fever, hypertension, heavy exercise, CCF
etc.
Post-renal proteinuria: caused by
imflammatory or neoplastic conditions in
renal pelvis, ureter, bladder, prostate or
urethra.
33. microalbuminuria
It is presence of albumin in urine above
normal level bt below detectable range of
conventional urine dipstick method.
Defined as urinary excretion of 30 to 300
mg/24 hrs of albumin in urine.
34. Significance of microalbuminuria
an indicator of subclinical cardiovascular
disease
an important prognostic marker for kidney
disease
in diabetes mellitus (earliest sign of renal
damage in DM)
in hypertension
increasing microalbuminuria during the first 48
hours after admission to an intensive care unit
predicts elevated risk for acute respiratory failure
, multiple organ failure , and overall mortality
35. Detection of microalbuminuria:
Methods for detection :
Measurement of albumin creatinine ratio
in random urine sample.
Measurement of albumin in early morning
sample.
Measurement of urine in 24 hr sample.
36. Bence Jones proteins
These are monoclonal immunoglobulin light
chains (kappa or lamda) synthesized by
neoplastic plasma cells.
seen in multiple myeloma, macroglobulinemias,
primary amyloidosis.
Test- Thermal method(waterbath):
Proteins have unusual property of
precipitating at 400
-600
c & then dissolving
when the urine is brought to boiling(1000
c) &
reappears when the urine is cooled.
37. Test for sugar Test-BENEDICT’S TEST(semiquantitative)
Principle-benedict’s reagent contains
cuso4.In the presence of reducing sugars
cupric ions are converted to cuprous oxide
which is hastened by heating, to give the
color.
Method- take 5ml of benedict’s reagent in a
test tube, add 8drops of urine. Boil the
mixture.
Blue= negative
Yellow=+(<0.5%)
green=++(0.5-1%)
Yellow-orange=+++(1-2%)
Brick red=++++(>2%)
38.
39.
40.
41. Benedict’s test
Detects all reducing substances like
glucose, fructose, & other reducing
sustances.
Sensitivity of the test is about 200 mg
reducing substance per dl of urine.
To confirm it is glucose, dipsticks can be
used (glucose oxidase)
42. Reagent strip method:
Specific for glucose.
Based on glucose oxidase peroxidase
reaction.
More sensitive (sensitivity- 100 mg
glucose/dl)
Glu + oxygen---- gluconic
acid+hydrogen peroxide
Hydrogen peroxide+chromogen--
oxidised chromogen(blue)+ H2O
44. Causes of glycosuria
Glycosuria with hyperglycaemia-
diabetes,acromegaly, cushing’s disease,
hyperthyroidism, drugs like corticosteroids.
Glycosuria without hyperglycaemia-
renal tubular dysfunction
45. Ketone bodies
3 types
Acetone
Acetoacetic acid
β-hydroxy butyric acid
They are products of fat metabolism
46. Rothera’s test
Principle-acetone & acetoacetic acid react
with sodium nitroprusside in the presence
of alkali to produce purple colour.
Method- take 5ml of urine in a test tube &
saturate it with ammonium sulphate. Then
add one crystal of sodium nitroprusside.
Then slowly run the liquor ammonia along
the sides of the test tube.
Formation of purple coloured ring at
junction indicates + test
47. Other tests:
Acetest tablet test
Ferric chloride test(Gerhardt’s test)
Reagent strip method
Hart’s test for beta hydroxy butyric acid
48. Causes of ketonuria
Diabetes
Non-diabetic causes- high fever,
starvation, severe vomiting/diarrhoea
Glycogen storage diseases
49. Bilirubin
Test- fouchet’s test.
In 5 ml of urine add 2.5 ml of 10% Barium
chloride and mix well. Then filter to obtain
precipitate. To ppt add 1 drop of fouchet’s
reagent. Development of blue green colour
indicate + test.
Principle: Bilirubin Adsorbs to the Barium Chloride
and results in green color formation when
fouchet’s reagent is added.
Causes
Liver diseases-injury,hepatitis
Obstruction to biliary tract
50. Other tests:
Foam test
Gmelin’s test (nitric acid)
Lugol’s iodine test
Reagent strip with diazo reagent
51. Urobilinogen
Test- ehrlich test
In 5 ml of urine add 0.5 ml of Ehrlich’s
reagent(HCl 20 ml, d/w 80 ml, p-
dimethylaminobenzaldehyde 2 gm). Allow
to stand for 5 min. development of pink
colour indicates +test.
Causes-hemolytic jaundice, early
hepatitis, hepatocellular jaundice.
52. Bile salt:
Hay’s sulphur test:
In 5 ml of urine sprinkle a pinch of
sulphur particles. If bile salt is present
sulphur particles will sink to the bottom
because bile salts lowers the surface
tension of urine.
53. Blood in urine
Test- BENZIDINE TEST
Principle-The peroxidase activity of hemoglobin
decomposes hydrogen peroxide releasing
nascent oxygen which in turn oxidizes benzidine
to give blue color.
Method- mix 2ml of benzidine solution with 2ml
of hydrogen peroxide in a test tube. Take 2ml of
urine & add 2ml of above mixture. A blue or
green color within 5 min indicates + reaction.
54. Causes of hematuria
Pre renal- bleeding diathesis,
hemoglobinopathies, malignant
hypertension.
Renal- trauma, calculi, acute & chronic
glomerulonephritis, renal TB, renal
tumors
Post renal – severe UTI, calculi,
trauma, tumors of urinary tract
55. Type Plasma color Urine color
Hematuria normal Smoky red
m/s-plenty of
RBC’s
hemoglobunuria Pink,hepatoglob
in reduced
Red ,
occasional
RBC’s
Myoglobunuria Pink, normal
hepatoglobin
Red, occasional
RBC’s
56. Microscopic examination
A well mixed sample of urine (12 ml) is
centrifuged in machine for 5 min at 1500 rpm.
The top liquid part (the supernatant) is
discarded. A drop of urine left at the bottom of
the test tube (the urine sediment) is placed on
glass slide and covered with cover slip. It is
examined under high power.
57. Contents of normal urine m/s
Contains few epithelial cells, occasional
RBC’s, few crystals.
70. casts
Urinary casts are cylindrical aggregations
of particles that form in the distal nephron,
dislodge, and pass into the urine. In
urinalysis they indicate kidney disease.
They form via precipitation of
Tamm-Horsfall mucoprotein which is
secreted by renal tubule cells.
72. Hyaline casts
The most common type of cast, hyaline
casts are solidified Tamm-Horsfall
mucoprotein secreted from the tubular
epithelial cells
Seen in fever, strenuous exercise,
damage to the glomerular capillary
74. Granular casts
Granular casts can result either from the
breakdown of cellular casts or the
inclusion of aggregates of plasma proteins
(e.g., albumin) or immunoglobulin light
chains
indicative of chronic renal disease
80. Fatty casts
Formed by the breakdown of lipid-rich
epithelial cells, these are hyaline casts
with fat globule inclusions
They can be present in various disorders,
including
nephrotic syndrome,
diabetic or lupus nephropathy,
Acute tubular necrosis
83. Pigment casts
Formed by the adhesion of metabolic
breakdown products or drug pigments
Pigments include those produced
endogenously, such as
hemoglobin in hemolytic anemia,
myoglobin in rhabdomyolysis, and
bilirubin in liver disease.
84. Crystal casts
Though crystallized urinary solutes, such
as oxalates, urates, or sulfonamides, may
become enmeshed within a hyaline cast
during its formation.
The clinical significance of this occurrence
is not felt to be great.
85. Red cell 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.
90. Epithelial casts
This cast is formed by inclusion or
adhesion of desquamated epithelial cells
of the tubule lining.
These can be seen in
acute tubular necrosis and
toxic ingestion, such as from mercury,
diethylene glycol, or salicylate.
100. Urine dipsticks
Urine dipstick is a narrow plastic strip which
has several squares of different colors
attached to it. Each small square represents
a component of the test used to interpret
urinalysis. The entire strip is dipped in the
urine sample and color changes in each
square are noted. The color change takes
place within 2 minutes from dipping the
strip. If read too early or too long after the
strip is dipped, the results may not be
accurate.
101. Chemical AnalysisChemical Analysis
Urine DipstickUrine Dipstick
Glucose
Bilirubin
Ketones
Specific Gravity
Blood
pH
Protein
Urobilinogen
Nitrite
Leukocyte Esterase
103. The squares on the dipstick represent the
following components in the urine:
∀ specific gravity (concentration of urine),
∀ acidity of the urine (pH),
∀ protein in the urine (mainly albumin),
∀ glucose (sugar),
∀ ketones
∀ blood
∀ bilirubin and
Urobilinogen
Nitrites
Leukocyte esterase
104. The main advantage of dipsticks is that
they are
1. convenient,
2. easy to interpret,
3. and cost-effective
105. The main disadvantage is that the
1. Information may not be very accurate as
the test is time-sensitive.
2. It also provides limited information about
the urine as it is qualitative test and not a
quantitative test (for example, it does not
give a precise measure of the quantity of
abnormality).
106. Automated urine analyser:
It is based on the principle of flow
cytometry.
This system classifies particles based on
fluoroscent intensity ( cells are stained
with 2 fluoroscent dyes), electrical
impedance and forward angle light scatter.
Next generation automated image based
urinalysis system iQ200 is a walk away
system.
107. This system uses flow imaging analysis
technology and Auto Particle Recognition
software.
It requires minimum of 3 ml of urine and
quantitates particles in 2 microlitre of
sample.
Can analyse 60 urine samples per hr.
108.
109. References:
Henry’s Clinical diagnosis and
Management of Laboratory Methods
Clinical Pathology Kawthalkar
Clinical Pathology Sabitri Sanyal
Basics of Body fluids by Akhil Bansal
Internet
Notas del editor
May occur singly or in clumps.
Mostly neutrophils, with granules n lobulation of nuclei.
&lt;5 leucocytes/hpf are seen normally.
Increased number of clumps are seen in cystitis, prostatitis and balanitis.
&gt;30/hpf suggests acute infection.
Eosinophils are increased in urine in tubulointerstitial disease, drug hypersesitivity.
May occur singly or in clumps.
Mostly neutrophils, with granules n lobulation of nuclei.
&lt;5 leucocytes/hpf are seen normally.
Increased number of clumps are seen in cystitis, prostatitis and balanitis.
&gt;30/hpf suggests acute infection.
Eosinophils are increased in urine in tubulointerstitial disease, drug hypersesitivity.
Under phase contrast, Trichomonas organisms appear much darker than the surrounding white blood cells.
This parasite is considered an important factor in the etiology of carcinoma of the bladder. The ova are elongated and are 60 X 160 microns. They are a yellowish color, slightly transparent and possess a delicate terminal spine.
Sperm may be present in urine sediment. Sperm have a characteristic oval body with a long thin tail and are 50 microns in length.
Oval fat bodies are degenerating tubular epithelial cells.
They contain refractile fat droplets.
These fats have been absorbed by the tubular cells after being leaked through abnormal glomeruli.
They appear as grape-like clusters of variable size and are highly refractile.
Yeast can appear as single cells or in the budding form.
In the budding form, yeast is easily identified as demonstrated on this slide.
Yeast can be found in patients with cystitis due to yeast, usually candida, or as a vaginal contaminant from patient&apos;s with vaginal candidiasis.