2. Why urine analysis?
• To fine out metabolic or endocrine
disturbances of the body
• To detect intrinsic conditions that may affect
the urinary tract and kidneys
• It is one of the oldest laboratory procedures
used in the diagnosis of diseases
4. • The GFR in children : 60ml/min
• It means that 60ml of glomerular filtrate is
formed per minute at the bowman’s capsule.
• About 180 liters of filtrate is formed in 24 hours
and due to reabsorption (99%) only 1.5 to 1.8L of
urine is excreted in 24 hours
5. CLINICAL CONDITIONS RELATED TO
URINE EXCREATION
1) Oliguria – urine output 800 to 2000ml per day
inspite of normal fluid intake/day
<1ml/kg/hour in infants
<0.5ml/kg/hour in children
- seen in shock , acute kidney injury
6. 2) Anuria : complete suppression of urine
formation
• seen in renal failure
3) Polyuria : urine output > 5ml/kg/hour or >2
liter/day/m2
• seen in diabetes insipidus , diabetes mellitus ,
bartter syndrome
7. COMPOSITION OF NORMAL URINE
• Volume : 600 to 2000ml/24 hour, Average:
1.2L
• Specific gravity : 1.003 to 1.030
• Reaction: acidic (pH : 4.7 to 7.5)
• Total solids : 30-70g/L
8.
9. SPECIMEN COLLECTION :
Should be collected in clean and dry wide mouth
glass/plastic containers with screw tops.
In infants and younger children : disposable collection
apparatus
• Time of urine specimen collection :
1. First void mid stream morning urine
2. The random sample
3. 24 hours urine specimen
10. METHODS OF URINE COLLECTION
1. Mid stream specimen : collected after voiding
the initial half of urine.
2. Clean catch specimen : urethral opening
cleaned with soap and water.
3. Catheter specimen : for bedridden , ill
patients or patients with obstruction of
urinary tract.
11. 4) Plastic bag specimen : in infants, clean plastic
bag is attached around the baby’s genitalia.
5) Suprapubic aspiration : used in infants
Urine aspirated from bladder by passing a needle
just above the symphysis pubis.
12.
13. PHYSICAL EXAMINATION
1. Volume : 1.2 to 1.5L /day
1. Color : Color of normal urine may vary from pale
yellow , dark amber depending upon the
concentration of pigments (urochrome) and
urobilinogen and uroerythrin.
1. Appearance : normal : clear.
• Cloudy urine : seen if amorphous phosphates present
in alkaline urine or amorphous ureate in acidic urine.
14. • Urine may appear cloudy or Turbid due to presence of
WBCs , bacteria and epithelial cells.
• Milky : due to presence of chile and fat
• Turbid and smoky : due to presence of RBCs
4 . Odor :
• Fruity odor : presence of ketone bodies .
• Pungent odor : Bacteria contaminated with urine due
to formation of ammonia.
• Musty odor : Urine in infant with phenyl ketonuria .
• Maple syrup like : maple syrup urine disease.
15. 5 . Reaction and PH:
• slightly acidic , normal PH: 4.6 to 7.
• Increase protein intake , ingestion of acidic fruits,
fever , starvation, respiratory acidosis and metabolic
acidosis and urinary tract infection produces acidic
urine .
• Diet rich in vegetables, citrus fruits, vomiting ,
respiratory alkalosis, metabolic alkalosis UTI caused by
proteus or ammonia produces alkaline urine.
16. 6. Specific gravity :
• Ratio of weight of a volume of urine to weight of
same volume of distilled water.
• Significance: used to measure the concentrating
power and diluting power of kidneys.
• Normal: 1.003 to 1.035(random sample)
1.015 to 1.030 (24 hour urine sample)
17. Isosthenuria : excretion of urine of fixed specific gravity of
1.010
which indicates poor tubular reabsorption.
Hyposthenuria : urine with low specific gravity.
seen in pyelonephritis, protien malnutrition, diabetes
insipidus, Renal tubular damage.
Hypersthenuria : excretion of urine with high specific gravity.
seen in low water intake , diabetes mellitus , albuminurea.
18. HOW DO WE MEASURE SPECIFIC GRAVITY
OF URINE ?
1. Urinometer – Bulb shaped instrument.
• Instrument is floated in a cylindrical container
containing urine.
• The depth to which it sinks in the urine
indicates the specific gravity.
2 . Urine strip method
3 . Refractometer
23. MOLISCH TEST • Monosaccharides when
treated with concentrated
sulphuric acid undergo
dehydration with the
removal of 3 molecules of
water. condense with
phenolic compounds to
give coloured products.
This forms the basis of
Molisch test. It is a
general test for
carbohydrates.
25. TESTS FOR LACTOSURIA
• Second most common
reducing sugar in urine.
• Helps to differentiate
between glucosuria and
lactosuria in Gestational
diabetes mellitus.
• METHYL AMINE TEST
• 5ml of urine+1ml of methyl
amine in sodium
hydroxide,kept at 56°C for
30 min.
• Red color indicates
presence of lactose.
26. TESTS FOR GALACTOSUREA
●Absence of galactose- 1-
phosphate uridyl
transferase.
MUCIC ACID TEST will be
positive. Boiling the urine
with nitric acid will lead to
formation of crystals of
mucic acid .
lactose or galactose is
present.
27. PENTOSUREA
●Deficiency of any of the
two enzymes xylitol
dehydrogenase or xylulose
reductase.
●Excretion of L-xylulose in
the urine.
●Bial's test will be positive.
The reagent contains
orcinol in HCl. 0.5 ml of
urine and 5 ml of Bial's
reagent are heated to boil. A
green color indicates the
presence of pentoses.
28. TESTS FOR PROTEINUREA
• Heat and Coagulation test.
• Take 10 mL of urine in a
test tube and boil the upper
part of the tube. If turbidity
appears, add six drops of
5% acetic acid. If turbidity is
due to the pres- ence of
phosphates, it will
disappear. If it persists, it
indicates that proteins are
present The presence of
protein is graded as 1 + to
4+
29. Observation Proteinurea Urinary protein
(mg/dl)
Slight turbidity-
letters can be read
1+ 15-30
More turbid 2+ 30-150
Cloudiness with fine
precipitate
3+ 150-300
More cloudy 4+ >300
30. SULPHOSALICYLIC ACID TEST
Add 5-10 drops of 20%
sulphosalicylic acid to 5
mL of urine and examine
for turbidity. Presence of
turbidity indicates
proteinuria and is graded
according to the
increasing amounts of
turbidity as l+ to 4+
31. Amount of turbidity Protein estimation
No turbidity Negative
Slight turbidity Trace(20mg/dl)
Turbidity is distinct
but without granular
formation
1+(50mg/dl)
Turbidity is distinct
and regular
2+(200mg/dl)
Dense turbidity with
distinct clumping
3+(500mg/dl)
Dense turbidity with
solidified large
clumps(precipitated
protein)
4+(1000mg/dl)or
more
32. Dip stick methods
• Widely used, more convenient and reliable.
• Light chain proteins, globulin and low
molecular weight tubular proteins are not
detected by this method.
• False negative result: dilute urine
• False positive results : alkaline urine,
concentrated urine , contamination with
chlorhexidine or benzalkonium.
34. • TAM HORSFALL PROTEIN
o it is glycoprotein from the ascending loop of
henley and epithelial cells of DCT.
o Also known as uromodulin -85 to 90 kd
glycoprotein.
o Normal excretion : 20 to 70 g/dl
35. ESBACH’S ALBUMINOMETER
Protein excretion in a 24-hour urine sample is required in
suspected cases of nephrotic syndrome (>3.5 g/24 hours)
and orthostatic/postural proteinuria.
Principle: Cold precipitation of proteins by a strong acid.
Procedure: Fill the albuminometer with urine up to the
mark U. Add Esbach’s reagent up to the mark R. Stopper
the Esbach’s albuminometer, mix and allow it to stand for
24 hours. Take the reading from the level of precipitation
in the albuminometer and divide the value by 10 to get
the percentage of total proteins.
36.
37. • ESTIMATION OF PROTEIN CREATININE
RATIO.
• Normal-<0.1
• Low to moderate proteinuria-0.1-2
• Nephrotic range(heavy) proteinuria->2
38.
39.
40. TESTS FOR KETONE BODIES
ROTHERA TEST
Saturate 5 ml of urine with
solid ammonium sulfate. Add 3
drops of freshly prepared
sodium nitroprusside followed
by 2 ml of liquor ammonia
along the sides of the test
tube. Development of a purple
ring at the junction of the two
liquids indicates the presence
of acetone or acetoacetic acid
in urine. It is not answered by
beta hydroxy butyrate.
Strip tests are also available.
41. • GERHADT’S TEST.
• To 5 ml of urine, add dilute
ferric chloride solution drop
by drop, till a maximum
precipitate of ferric
phosphate is obtained. This
is to eliminate the
phosphates which may
obscure the color in the
test. Filter. To the filtrate
add excess ferric chloride. A
red color indicates the
presence of acetoacetic
acid.
42. Causes for Ketone bodies.
Diabetic ketonuria
• Diabetic ketoacidosis
Nondiabetic ketonuria
• Starvation
• Prolonged vomiting or diarrhea
• Infant and children – Prolonged febrile illness – Toxic states
accompanied by vomiting or diarrhea – Glycogen storage
disorders (von Gierke’s disease)
Lactic acidosis
• Shock, diabetes mellitus, renal failure, liver disease,
infection, and drugs (e.g. phenformin and salicylate
poisoning)
43. TESTS FOR BILIRUBIN
• FOUCHET’S TEST
• Take 10 ml of urine+2 ml
of barium chloride.
• Filtrate the urine,filtrate
is put on the filter paper.
• Add one drop of ferric
chloride.
• Green colour indicates
,Bilverdin is present,
which is formed from
Bilirubin.
44. TEST FOR UROBILINOGEN
• EHRLICH’S TEST
• 10ml of urine+1ml of
Ehrlich’s benzaldehyde
reagent.
• Mix it and keep for 10
minutes.
• Development of pink
color indicates normal
amount of
bilinogen.Darkred color
means increased amount.
45. TEST FOR BILE SALTS
• HAY’S TEST
• 5ml of urine
• Sprinkle finely
powdered Sulphur on
the surface of urine.
• Sulphur will sink down
indicating the presence
of BILE salts.
46. URINARY FINDINGS IN JAUNDICE
TYPE OF JAUNDICE BILE PIGMENT BILE SALT UROBILINOGEN
PREHEPATIC
(HEMOLYTIC)
Nil Nil ++
HEPATOCELLULAR ++ + Normal or
decreased
POST HEPATIC
(OBSTRUCTIVE)
+++ ++ Nil or decreased
47. MICROSCOPIC EXAMINATION OF
URINE
A fresh well mixed specimen is examined for
cellular elements, crystals and casts.Urine is
centrifuged at 1500rpm for 10min,urine is
decanthed and the cell pellet resuspended in 0.3 -
0.5 ml of urine.
Neutrophils nil
Erythrocytes(RBC) 0-4 HPF
Epithelial cells 0-5HPF
Casts nil
Crystals nil
51. TESTS FOR HEMATURIA
BENZIDINE TEST
• Make saturated solution of
benzidine in glacial acetic
acid.
• Mix 1 ml of this
solution+1ml of hydrogen
peroxide+2ml of urine.
• If green color or blue color
develops within 5 minutes
the test is positive.
ORTHOTOLUIDINE TEST
• Instead of benzidine
,orthotoludine is used.
• More sensitive than
benzidine test.
63. URINE CULTURE
Collection of urine is best done by
percutaneous suprapubic bladder puncture.
Significant bacteriuria refers to colony count
of more than 100,000 organisms per millilitre
and is seen in urinary tract infections.
64. • E. coli( uropathoenic E. coli or UPEC) is the
single most common
pathogen, accounting for 70-75% of all cases of
UTI.UPEC serotype
01, 02, 04, 06, 07 and 075 are responsible for
most UT!s.
65. • E. coli is an aerobe and facullative anaerobe.
• lt grows on ordinary culture media at optimum
temperature of 37•c (ranges 10-40'C) in 18-24 hours.
• Blood agar: Colonies are big,circular, grey, moist and
occasionally hemolytic (mainly UPEC strains).
• MacConkey agar: Colonies are circular, moist,
smooth with entire margin, flat and pink (due to
lactose fermentation).
• Peptone water broth produces uniform turbidity.
