Urine Analysis Practical

8/12/2019 Urine Analysis Practical

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Urine Analysis


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Urine Constituents In general , urine consists of urea and other

organic and inorganic chemicals dissolved inwater

Urea accounts for almost half of the totaldissolved solids in urine

Organic substances include creatinine and uricacid

The major inorganic solid dissolved in urine ischloride, followed by sodiumand potassium

Other substances found in urine includehormones , vitaminsandmedications

Other formed elements as cells ,casts, crystals,mucusand bacteria.


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Collection of Urine

Random Sample :

Which is sutable for routine

physical ,chemical andmicroscopic examinations

First Morning Sample :

The most prefered sample for

routine urine analysis because it

is the most concentrated sample


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Urine Constituents

Factors affecting urine constituents :

Dietary intake

Physical activity

Body metabolism

Kidney function

Endocrine function

Water intake

Drugs

Pregnancy stress


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Collection of Urine

Timed Sample :

12 hours timed sample is

required to clearance test orenzyme excretion tests.

24 hours timed sample isrequired for quentitation of

substance as hormones

,protein and electrolytes


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Urinary Catheter
http://www.le-west.co.uk/photos/M0291-1-F.JPG


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Urinary Collection Bag


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Storage of Urine Samples

Instructions

1.Urine should be either examined within 2-3

hours of collection or preserved in some

manner, usually by refrigeration (2 to 8 oC).

2.Appropriate preservatives may also be

used .

3.Specimen containers should be marked

with the

patients date and time of collection.


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Handling of Urine Samples

1. Centrifuge a 10 mL aliquot of urine for 10

minutes at 400g.2. Remove 9.5 mL of the supernatant urine.

3.Resuspend the sediment gently but

thoroughly in the remaining 0.5 mL of urineby a pipette to a glass slide covered with a

32X24 mm coverslip .

4.Examine the urine sample at low (160X) andhigh (400X) magnification within 2-3 hours

of collection.

5.Particles are expressed as lowest and


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Handling of Urine Samples

.

Centrifugation Discard 9.5 ml Resuspend

10 mL urine

sediment

supernatant


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Urinalysis

.

A. Physical Examination

Includes:

1. Volume.

2. Color.

3. Odor.4. Aspect.

5. Reaction (pH).

6. Specific gravity.B. Biochemical Examination

Includes:1. Proteins.

2. Sugers.

3. Ketone bodies.

4. Bile salts.

5. Bile Pigments.

6. Blood.

C. MicroscopicTestsInclude:

1. Cells.

2. Crystals.

3. Casts.4. Ova.

5. Parasites.


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Physical Examination of Urine

This includes

1.Volume.2.Color.

3.Odor.

4.Aspect.

5.Reaction.

6.Specific gravity.


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Volume of Urine

Normal volume:600-2500 ml/24 hrs.

Abnormalities of urine volume

a. Polyuria: abnormal increase in

urine volume. (>2500ml/24hrs) e.g. diabetesinsipidus (deficiency of ADH)

and diabetes mellitus(deficiency of insulin) . Chronicrenal failure and sever tubular


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Oliguria: abnormal decrease in

urine volume (< 500ml/24hrs) e.g. shock and acutenephritis dehydration renal

ischaemia as in heart failure andshock , chronic nephritis ,acutetubular necrosis ,acuteglomerulonephritis andobstructionc. Anuria: urine volume is lessthan 1oo ml, complete

suppression of urine formation


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Color of UrineNormal color: light amber color due to

urochrom pigment and small amountsof urobilin and uroerythrine .

Abnormal colors :

1. Greenish yellow: due to bilirubin e.g.

jaundice

2. Black urine : in alkaptonuria ,methemoglobinurea ,melaninuria and

by some drugs

3. Milky urine : due to lymph or lipid


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Color of Urine

4. Red Urine :

On eating beets

On taking certain drugsHemoglobinuria ( bright red and clear )

Acute glomurlonephritis ( red and smoky

)Porphyrinuria ( red and purple )


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Turbidity

Turpid urine may be due to : Phosphate precipitation :

it is present in alkaline urine

It fades by addition of acetic acid or anyacid

It increase by heating

Urate preceptation : It is present in acid urine

It forms white or pink colour ppt

It disappears on heating


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Turbidity

Pus cells : form white ppt ,it doesnot

disappear on heating but it becomes

gelatenous and its ppresence is confirmed

by microscopic examination

Bacteria growth : gives uniform

cloudiness that cannot be removed by

filtration or centrifugation


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Turbidity

Mucus : it form bulky deposite and

disappears by addition of acetic acid

Red cells : gives turbid smoky urine

and confirmed by microscopic

examination

Chyluria : gives turbid milky urine ,

urine becomes clear when shaked with

an e ual volume of etheror chloroform


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Odor of Urine

Normal odor

Fresh urine has aromatic odor or urineferousodor due to presence of volatile acids

aromatic acids.

Abnormal odorsa. Ammonia smell: after prolonged standing

b. Fecal smell: due to urinary infection

c. Fruity smell: ketosis (e.g. due touncontrolled

diabetes mellitus)


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Urine Sample

CHEMSTIX ANALYSIS

We are now ready to perform tests on the urine

sample that will analyze for the followingcharacteristics:

-specific gravity

-pH-protein

-glucose

-ketones

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-nitrites

-bilirubin-hematuria

-urobilinogen

-leukocytes

Chemstix Urinalysis

Data Sheet
http://localhost/var/www/apps/conversion/tmp/scratch_10/CHEMSTIX%20URINALYSIS.pdfhttp://localhost/var/www/apps/conversion/tmp/scratch_10/CHEMSTIX%20URINALYSIS.pdfhttp://localhost/var/www/apps/conversion/tmp/scratch_10/CHEMSTIX%20URINALYSIS.pdf


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Urine Sample

SPECIFIC GRAVITY

Normal specific gravity of urine is measured at

1015

1025.

Abnormally low specific gravity

(hyposthenuria : sg 1001-1010 ) indicates

dilute urine, which may be caused by:-Diabetes insipidus

- drinking excessive amounts of liquid

- severe kidney disease- the use of diuretics

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SPECIFIC GRAVITY


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SPECIFIC GRAVITY

Abnormally high specific gravity

(hypersthenuria :sg 10251035 ) indicates

very concentrated urine, which may becaused by:

- not drinking enough liquid

-loss of too much liquid (excessive

vomiting, sweating, or diarrhea)

-Diabetes mellitus( sugar )

- Toximea of pregnancy (protein in the

urine)


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Specific gravitiy measurement

1.refractometer(total solids meter ):

It measure the ratio of the velocity of light

in air to the velocity of light in urine

2. urinometer: less accurate ,the

urinometer is floated in the urine ,the sg is

read off the urineometer at the meniscus

level of the urine

3. the multiple test dipstick : an indicator

changes color in relation to ionic

concentration


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Urine Sample

pH

Normal pH for urine ranges from 4.5 8.0.

Some foods (such as citrus fruit and dairy

products) and medications (such as antacids)

can affect urine pH. In a diet high in protein the

urine is more acidic, while a diet high in

vegetable material yields a urine that is morealkaline.

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Urine Sample

pH

A high (alkaline) pH can be caused by

prolonged vomiting, a kidney disease, some

urinary tract infections, and asthma.

A low (acidic) pH may be a sign of severe lung

disease (emphysema), uncontrolled diabetes,aspirin overdose, prolonged diarrhea,

dehydration, starvation, drinking an excessive

amount of alcohol, or drinking antifreeze

(ethylene glycol).

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Urine Sample

PROTEIN

Proteinuria is usually a sign of kidney

disorders, but it may occur normally after

strenuous exercise such as marathon

running. Fever, strenuous exercise, normal

pregnancy, and some diseases (especially

kidney disease) may also cause protein inthe urine. Protein in the urine can also be

caused by heart failure, leukemia, poison

(lead or mercury poisoning), or a condition

during pregnancy that results in high bloodpressure (preeclampsia).

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P t i i U i (P t i i )


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Proteins in Urine (Proteinuria)

Types

I. Albuminuria- Normally traces of albumin are excreted inurine

daily.

- Causes of pathological albuminuriaa- Pre-renal

The primary causes are factors operating

beforethe kidney e.g. heart failure.

b- Renal

The lesion is intrinsic in the kidney e.g.


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II. Bence-Jones Proteins

- These are abnormal globulins that appear in

urine

in the following conditions

a. Multiple myeloma: a bone marrow cancer.

b. Leukemia.

- Urine in such conditions undergo 3 phases;

1.Clotting when heated to 60o

C, undergoes2.Dissolution of the clots when boiled at 100

C0.

3.Re-clotting upon cooling.

Gl i U i (Gl i )


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Glucose in Urine (Glucosuria)

Causes

1.Diabetes mellitus.2.Epinephrine glucosuria due to stress,

emotion,

fear, or anger.3.Renal glucosuria due to:

a.Congenital defects in renal tubular re-

absorption

of glucose.

b.Nephritis.


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Urine Sample

KETONES

Large amounts of ketones in the urine may

signal a dangerous condition known asdiabetic ketoacidosis. Ketones in the urine

can indicate poorly controlled diabetes, a

very low-carbohydrate diet, starvation

(including disorders that result in poor

nutrition such as anorexia nervosa or

bulimia), alcoholism, or poisoning from

drinking rubbing alcohol (isopropanol).

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Urine Sample

NITRITES

A positive nitrite test indicates that bacteria

may be present in significant numbers in

urine. Gram negative rods such as E. coli

are more likely to give a positive test. High

nitrite levels indicate an infection.

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Urine Sample

BLOOD

This test is based on detection of the

molecules of heme (present in hemoglobinor myoglobin). Blood in the urine

(hematuria) is detectable by Chemstix and

confirmed by viewing the urine with a

microscope. Sometimes the urine contains

enough blood to be visible, making the

urine appear red or brown.

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Urine Sample

BILIRUBIN

The color change indicating a positive

reaction, however, is a rather subtletransition among shades of beige, and

sometimes is obscured by color inherent in

the urine itself. Detection of bilirubin in

urine is generally an abnormal finding.

Bilirubinuria generally results when

conjugated bilirubin levels in blood are

elevated as a result of hepatobiliarydisease.

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Urine Sample

LEUKOCYTE

Leukocyte esterase (an enzyme found in

certain white blood cells) in the urine can bedetected by Chemstix. Leukocyte esterase

is a sign of inflammation, which is most

commonly caused by a urinary tract

infection. A positive leukocyte esterase test

results from the presence of white blood

cells either as whole cells or as lysed cells.

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Urine Sample

LEUKOCYTE

A negative leukocyte esterase test means

that an infection is unlikely and that,without additional evidence of urinary tract

infection, microscopic exam and/or urine

culture need not be done to rule out

significant bacterial infection in the urinary

tract.

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Urine Sample

UROBILINOGEN

Normal ranges of urobilinogen are 0.2 to 1.

Increases in the secretion of urobilinogenindicate significant hemolysis of

erythrocytes to the point that the liver

cannot process the bilirubin. The bilirubinincreases in the plasma and the formation

of urobilinogen in the intestines increases

as well. The urobilinogen diffuses into theblood, where it is filtered by the kidneys.

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Urine Sample

MICROSCOPIC ANALYSIS

Sediment in urine can be examined under amicroscope to provide information about a

possible kidney or urinary tract disorder.

Normally, urine contains a small number of

cells and other debris shed from the inside

of the urinary tract. A person who has a

kidney or urinary tract disorder usually

sheds more cells, which form a sediment ifurine is centrifuged or allowed to settle.

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Urine Sample

MICROSCOPIC ANALYSISIn this test, urine is spun in a centrifuge so

the solid materials (sediment) settle out.

The sediment is spread on a slide and

examined under a microscope.

Types of materials that may be found

include:-Microorganisms

-Cells

-Crystals

-Casts and FibersBe sure you have printed off the Data Sheet

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Microscopic Observations

Data Sheet
http://localhost/var/www/apps/conversion/tmp/scratch_10/MICROSCOPIC%20OBSERVATIONS.pdfhttp://localhost/var/www/apps/conversion/tmp/scratch_10/MICROSCOPIC%20OBSERVATIONS.pdfhttp://localhost/var/www/apps/conversion/tmp/scratch_10/MICROSCOPIC%20OBSERVATIONS.pdf


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Urine Sample

MICROSCOPIC ANALYSIS

In the sample you have looked at under the

microscope, we will attempt to quantify andidentify if the following are present in our

sediment sample:

-Red blood cells-White blood cells

-Casts

-Epithelial cells

-Bacteria-Crystals

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Urine Sample

RED BLOOD CELLS

Red blood cells are reported quantitatively

as number seen per Highpower field (HPF):none seen

100

Red blood cells are normal in urine in low

numbers. Up to 5 RBC/HPF generally are

considered acceptable.

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Urine Sample

RED BLOOD CELLS

Hematuria is the presence of abnormal

numbers of red blood cells in urine due toglomerular damage, kidney trauma, urinary

tract stones, urinary tract infections, blood

toxins, and physical stress. Red cells may

also contaminate the urine from the vagina

in menstruating women. Theoretically, no

red blood cells should be found, but some

find their way into the urine even in veryhealthy individuals.

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Urine Sample

WHITE BLOOD CELLS

Pyuria refers to the presence of abnormal

numbers of WBC that may appear withinfection in the urinary tract. WBC from the

vagina, especially in the presence of

vaginal and cervical infections, or the

urethra in men and women maycontaminate the urine.

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WBC

RBC


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Urine Sample

CASTSSome types of kidney disease can

cause plugs of material (called casts) to

form in the nephrons of the kidneys.

Casts are cylindrical protein-based

molds of the nephron tubule which can

then get flushed out into the urine.

Casts can be made of different types of

material, such as red or white blood

cells, waxy or fatty substances, or

protein. The type and make-up of cast

can provide clues about the type of

kidney disease that may be present.

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Urine Sample

CASTS

Depending on the type, casts can indicate

inflammation or damage to the nephrons inthe kidneys, poor blood supply to the

kidneys, metal poisoning (such as lead or

mercury), heart failure, or a bacterial

infection.They are absent or very few inurine samples.The numbers of casts seen

are usually reported as number of each type

found per Lowpower field (LPF).

Example: 5-10 casts/LPF.

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Urine Sample

CASTS

Depending on the type, casts can indicate

inflammation or damage to the nephrons inthe kidneys, poor blood supply to the

kidneys, metal poisoning (such as lead or

mercury), heart failure, or a bacterial

infection.They are absent or very few inurine samples.The numbers of casts seen

are usually reported as number of each type

found per Lowpower field (LPF).

Example: 5-10 casts/LPF.

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Urine Sample

CRYSTALS

Most often, crystals in routine urine

sediment preps are without significance.Several different types can be seen in

normal samples. A few specific types,

however, can be important in certain clinical

situations. Shown below are struvitecrystals (magnesium ammonium

phosphate), which are a common finding in

normal urine.

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Urine Sample

MICROORGANISMS

Bacteria can be common in urine

specimens because of the abundant normalmicrobial flora of the vagina or male urethra

and because of their ability to rapidly

multiply in urine standing at room

temperature. Therefore, microbialorganisms found in all but the most

scrupulously collected urines should be

interpreted in view of clinical symptoms.

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Urine Sample

MICROORGANISMS

Diagnosis of bacteriuria in a case of

suspected urinary tract infection requiresculture. A colony count may also be done to

see if significant numbers of bacteria are

present. Generally, more than 100,000/ml of

one organism reflects significantbacteriuria.

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C S


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Urine Sample

EPITHELIAL CELLS

Epithelial cells in urine are generally of little

specific diagnostic utility. Cells lining theurinary tract at any level may slough into

the urine. In the case of voided samples,

even cells from the genital tract can appear

in the sample. Most commonly seen areepithelial cells from the urethra, vulva,

bladder and urethra.

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CANCER CELLS


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Urine Sample

CANCER CELLS

Microscopic examination of the urine to

look for cancer cells, is sometimes useful indiagnosing cancers of the kidneys and

urinary tract. For people at high risk (for

example, smokers, petrochemical workers,

and people with painless bleeding) urinesediment analysis may be used to screen

for cancer of the bladder and kidneys.

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Urine Analysis Practical

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