URINE ANALYSISURINE ANALYSIS

1.Formation of Urine1.Formation of Urine

2.Normal constituents and composition2.Normal constituents and composition

3.Collection of urine specimens3.Collection of urine specimens

4.Preservation of urine4.Preservation of urine

5.Physical examination of urine5.Physical examination of urine

6.Chemical Examination of Urine 6.Chemical Examination of Urine

7. Microscopic Examination Of Urine Deposits7. Microscopic Examination Of Urine Deposits

• Urine --- most easily obtainedUrine --- most easily obtained

• Examination of urine Examination of urine

• Information--- about the functioning Information--- about the functioning of the kidney of the kidney

• diagnosis ---of urinary tract diseases diagnosis ---of urinary tract diseases

• diagnosis -----of certain metabolic diagnosis -----of certain metabolic and systemic diseases. and systemic diseases.

• Urine is formed in the kidney. Urine is formed in the kidney.

• The functional unit of kidney is called The functional unit of kidney is called nephrons,nephrons,

• where in the ultrafilteration of plasma takes where in the ultrafilteration of plasma takes place, place,

• followed by absorption of most of the water followed by absorption of most of the water and some of the solutes. and some of the solutes.

• The kidneys through the nephrons ---excrete The kidneys through the nephrons ---excrete many of the waste products of the bodymany of the waste products of the body

3 NORMAL CONSTITUENTS AND 3 NORMAL CONSTITUENTS AND COMPOSITION OF URINECOMPOSITION OF URINEConstituent’s Constituent’s grams/litregrams/litreA.InorganicA.Inorganic• Chloride 9.0Chloride 9.0• Phosphorous 2.0Phosphorous 2.0• Sulfur 1.5Sulfur 1.5• Sodium 4.0Sodium 4.0• Potassium 2.0Potassium 2.0• Calcium 0.2Calcium 0.2• Magnesium 0.2Magnesium 0.2• Iron 003Iron 003B.OrganicB.Organic• Urea 25Urea 25• Uric acid 0.6Uric acid 0.6• Creatinine 1.5Creatinine 1.5• Ammonia 0.6Ammonia 0.6• Sugar not detected by benedicts test traceSugar not detected by benedicts test trace• Ketone bodies traceKetone bodies trace• Carbonates, bicarbonates & free carbonic acid traceCarbonates, bicarbonates & free carbonic acid trace• Mucin & mucin like substances DiastaseMucin & mucin like substances Diastase

COLLECTION OF URINE SPECIMENSCOLLECTION OF URINE SPECIMENS • In. Improper collection---- may invalidate the results In. Improper collection---- may invalidate the results • Containers for collection of urine should be wide Containers for collection of urine should be wide

mouthed, clean and drymouthed, clean and dry• First morning sampleFirst morning sample –concentrated urine --- –concentrated urine --- chemical constituents chemical constituents casts and crystals casts and crystals • Random specimenRandom specimen chemical screening chemical screening microscopic examinationsmicroscopic examinations• 24 hours urine sample24 hours urine sample quantitative estimation of proteins, sugars, quantitative estimation of proteins, sugars,

electrolytes, and hormones. electrolytes, and hormones. Urine specimen is collected in a clean 2 liters bottle with Urine specimen is collected in a clean 2 liters bottle with

a stopper. The first morning sample is not collected. a stopper. The first morning sample is not collected. All the urine passed during the rest of the day and night All the urine passed during the rest of the day and night

and next day I st sample is collected. and next day I st sample is collected. Volume is measured and immediately sent to the Volume is measured and immediately sent to the

laboratory.laboratory.

Midstream urine specimenMidstream urine specimen• Urine should be collected in a clean, dry Urine should be collected in a clean, dry

and, preferably, sterilized container. and, preferably, sterilized container. • urethral opening is cleaned with a moist urethral opening is cleaned with a moist

cotton swab. cotton swab. • first 10 – 25 ml of urine is not collected first 10 – 25 ml of urine is not collected

(discarded)(discarded)• since it contains urethral and prostatic since it contains urethral and prostatic

secretions which may be required if secretions which may be required if investigating urethra and prostrate. investigating urethra and prostrate.

• First morning samples ---- these are the First morning samples ---- these are the concentrated samples and casts, crystals concentrated samples and casts, crystals etc .if present etc .if present

• Terminal urine sample patient voids the last Terminal urine sample patient voids the last portion of urine in an open container.portion of urine in an open container.

• Urine specimen collection using a catheter. This Urine specimen collection using a catheter. This procedure is used for certain bacteriological tests procedure is used for certain bacteriological tests

• Urine specimens from infantsUrine specimens from infants• Urine can be collected into a plastic bag with an Urine can be collected into a plastic bag with an

adhesive mouth. adhesive mouth. • The bag is fixed around the genitalia and left The bag is fixed around the genitalia and left

in place for 1- 3 hours, depending on in place for 1- 3 hours, depending on the examination requested. the examination requested.

• Colostomy bags can be usedColostomy bags can be used

PRESERVATION OF URINEPRESERVATION OF URINE

• Urine sample < within 2 hours. Urine sample < within 2 hours.

• If delay ----preservation. If delay ----preservation.

• If more than 2 hoursIf more than 2 hours– Keep the sample in the refrigerator without any Keep the sample in the refrigerator without any

preservative.preservative.– Toluene – add a few drops till it forms a thin layer Toluene – add a few drops till it forms a thin layer

on the surface of urine.on the surface of urine.– Conc.HCL – 1 ml of conc.HCL for 125-150mlof urine.Conc.HCL – 1 ml of conc.HCL for 125-150mlof urine.– Formaldehyde -1 drop for 15 ml of urine. Cells and Formaldehyde -1 drop for 15 ml of urine. Cells and

casts are well preserved.casts are well preserved.

PHYSICAL EXAMINATIONPHYSICAL EXAMINATION

• VolumeVolume – – normal -- 1.2-2 L /day normal -- 1.2-2 L /day the day is 3-4 times > night. the day is 3-4 times > night. night is < 400 ml. night is < 400 ml. polyuria polyuria >2000ml / day. >2000ml / day. OliguriaOliguria <500ml / day. <500ml / day. AnuriaAnuria is total suppression of urine <100 ml per is total suppression of urine <100 ml per

day.day.• Appearance – color & turbidityAppearance – color & turbidity• 1. Color1. Color - normal ---- amber yellow ( to the - normal ---- amber yellow ( to the

presence of urobilin,uroerythrin,and urochromes)presence of urobilin,uroerythrin,and urochromes)

1.Colorless---Very dilute urine1.Colorless---Very dilute urine ---Polyuria,diabetes---Polyuria,diabetes 2.Yellow orange(high colored)--Concentrated urine2.Yellow orange(high colored)--Concentrated urine -----------Excess urobilin-----------Excess urobilin ------------Bile pigments------------Bile pigments ------------Intake of carrots ------------Intake of carrots (Yellow foam ++)(Yellow foam ++) 3.Red/ smoky------Hemoglobin/3.Red/ smoky------Hemoglobin/ ---- RBC---- RBC ----MyoglobinBeetroot /----MyoglobinBeetroot / ---------- anilinedyes ---------- anilinedyes -------Menstrual contamination-------Menstrual contamination -------porphyrins -------porphyrins 4.Cloudy--------Phosphates & carbonates, Urates & 4.Cloudy--------Phosphates & carbonates, Urates &

uric acid Pus cells uric acid Pus cells bacteria,yeast,spermatozoabacteria,yeast,spermatozoa 5.Milky-------Pyuria,Fat ,chyluria5.Milky-------Pyuria,Fat ,chyluria 6. Brown black---- 6. Brown black----

Methemoglobin,,Homogenesticacid(alkaptonuria),MeMethemoglobin,,Homogenesticacid(alkaptonuria),Melaninlanin

7. orange--------Bile pigments,Drugs – 7. orange--------Bile pigments,Drugs – pyridium,rifampicinpyridium,rifampicin

• 2. Turbidity2. Turbidity – normal urine----clear. – normal urine----clear.

• Turbidity in urine may be due to:Turbidity in urine may be due to:

a.a. amorphous phosphate and carbonates – alkaline amorphous phosphate and carbonates – alkaline or or

neutral urine neutral urine disappears on addition of dilute acetic acid disappears on addition of dilute acetic acid

b. crystals, cellular exudates, bacteria and fungusb. crystals, cellular exudates, bacteria and fungus

c. chyle and fat c. chyle and fat

d. pus d. pus

e. amorphous urates in acidic urinee. amorphous urates in acidic urine

• b. Odourb. Odour: Normal-- aromatic odour : Normal-- aromatic odour

On standing--- ammonical On standing--- ammonical

there is decomposition of urea there is decomposition of urea forming ammonia which gives a forming ammonia which gives a strong ammonical smell.. strong ammonical smell..

• Abnormal odour of urineAbnormal odour of urine

Fruity---- Ketonuria Fruity---- Ketonuria

Mousy---- PhenylketonuriaMousy---- Phenylketonuria

Rancid------ Tyrosinaemia Rancid------ Tyrosinaemia

pHpH•

ability to maintain normal hydrogen ion concentrationability to maintain normal hydrogen ion concentration • Normal kidneys are capable of producing urine the pH of Normal kidneys are capable of producing urine the pH of

which varies from 4.5 to slightly higher than 8.0. which varies from 4.5 to slightly higher than 8.0.

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

• PROCEDUREPROCEDURE

• Dip the litumus paper strips in the urine, remove and read Dip the litumus paper strips in the urine, remove and read the color change immediately.the color change immediately.

• Blue litmus turns red - acidBlue litmus turns red - acid• Red litmus turns blue - alkalineRed litmus turns blue - alkaline• Blue and red litmus turns purple - neutral Blue and red litmus turns purple - neutral • nitrazine paper method :The Ph---- nitrazine paper nitrazine paper method :The Ph---- nitrazine paper

which are sensitive and specific in the pH range of which are sensitive and specific in the pH range of 4.5 – 8.0 range. 4.5 – 8.0 range.

SPECIFIC GRAVITYSPECIFIC GRAVITY

• degree of concentration or dilution degree of concentration or dilution of the specimen. of the specimen.

• specific gravity measures the specific gravity measures the concentrating and diluting abilities concentrating and diluting abilities of the kidney. of the kidney.

• The normal --------------1.015 and The normal --------------1.015 and 1.025 (in a 24 hours specimen).1.025 (in a 24 hours specimen).

ESTIMATION OF SPECIFIC GRAVITY ESTIMATION OF SPECIFIC GRAVITY BY URINOMETERBY URINOMETER

• urinometer: This is a weighted bulb-urinometer: This is a weighted bulb-shaped shaped

a scale calibrated----- from 1.000 a scale calibrated----- from 1.000 to 1.060 to 1.060

PROCEDUREPROCEDUREUrine is poured into a cylindrical or conical glass so that the Urine is poured into a cylindrical or conical glass so that the

vessel is nearly full.vessel is nearly full. Froth-- removed with a filter Froth-- removed with a filter • The instrument is floated in the urine and care should be The instrument is floated in the urine and care should be

taken to see that it does not touch the slides. taken to see that it does not touch the slides.

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

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

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

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

equal volume of distilled water and the last two figures of equal volume of distilled water and the last two figures of the reading are then multiplied by 2.the reading are then multiplied by 2.

• Correction for temperatureCorrection for temperature- - • The urinometer is calibrated to read 1.000 in The urinometer is calibrated to read 1.000 in

distilled water at a specific temperature, distilled water at a specific temperature, indicated on each instrument e.g. 15 C or 20 C. indicated on each instrument e.g. 15 C or 20 C.

• There is a change in the specific gravity of 0.001 There is a change in the specific gravity of 0.001 for each 3 C above and below this temperature. for each 3 C above and below this temperature. Therefore add 0.001 to the reading for each 3 C Therefore add 0.001 to the reading for each 3 C above the temperature for which the urinometer above the temperature for which the urinometer is calibrated, is calibrated,

• substract 0.001 for each 3 C, the temperature substract 0.001 for each 3 C, the temperature below the standard temperature.below the standard temperature.

• For exampleFor example - urinometer calibrated for 20 C - urinometer calibrated for 20 C • Specific gravity of urine at 32 C is 1.001Specific gravity of urine at 32 C is 1.001• Corrected specific gravity {(32 – 20)/ 3 X 0.001} Corrected specific gravity {(32 – 20)/ 3 X 0.001}

+ 1.001 = 1.005+ 1.001 = 1.005• Correction is also recommended when glucose or Correction is also recommended when glucose or

protein are present. It is recommended that .003 protein are present. It is recommended that .003 be subtracted from the urinometer reading for be subtracted from the urinometer reading for each 1000 mg / dl of glucose or protein.each 1000 mg / dl of glucose or protein.

• 1. SUGARS IN URINE: 1. SUGARS IN URINE:

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

• Benedict’s qualitative test:Benedict’s qualitative test:

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

• Composition of Benedict’s reagent:Composition of Benedict’s reagent:

Copper sulphate - 17.3 gmsCopper sulphate - 17.3 gms Sodium carbonate (anhydrous) – 100 gms Sodium carbonate (anhydrous) – 100 gms OrOr Crystalline sodium carbonate - 200 GmsCrystalline sodium carbonate - 200 Gms Sodium citrate - 175 gmsSodium citrate - 175 gms OrOr Potassium citrate Potassium citrate

• Dissolve crystalline copper sulphate in 100 Dissolve crystalline copper sulphate in 100 ml of distilled water. Dissolve sodium ml of distilled water. Dissolve sodium carbonate and sodium citrate in 700 cc. of carbonate and sodium citrate in 700 cc. of distilled water. Slowly add the latter to the distilled water. Slowly add the latter to the former solution with constant stirring. When former solution with constant stirring. When complete, make up the volume to 1000ml complete, make up the volume to 1000ml with distilled water.with distilled water.

Procedure Procedure

• Take 5ml of benedicts reagent Take 5ml of benedicts reagent

• Boil for 3 – 5 minutesBoil for 3 – 5 minutes

• add to it 0.5ml (8 drops)of protein free urine.add to it 0.5ml (8 drops)of protein free urine.

• Cool and note the color. Cool and note the color.

Recording resultsRecording results

The color varies from blue through green – The color varies from blue through green – yellow- orange- brick red. yellow- orange- brick red.

Negative – no change in color.Negative – no change in color.

Trace - solution appears pale green to Trace - solution appears pale green to slightly cloudy.slightly cloudy.

1+ - Definite cloudy green (0.5% sugar)1+ - Definite cloudy green (0.5% sugar)

2+ - Yellow to orange precipitate, 2+ - Yellow to orange precipitate, supernatant fluid pale blue (1% sugar)supernatant fluid pale blue (1% sugar)

3+ - Orange to red precipitate, supernatant 3+ - Orange to red precipitate, supernatant fluid pale blue (1.5% sugars) fluid pale blue (1.5% sugars)

4+ - Brick red precipitate, supernatant fluid 4+ - Brick red precipitate, supernatant fluid decolorizes (2 % sugar) decolorizes (2 % sugar)

– COLORIMETRIC REAGENT STRIP TEST COLORIMETRIC REAGENT STRIP TEST

• Principle: this test is based on a Principle: this test is based on a double sequential enzyme reaction.double sequential enzyme reaction.

• One enzyme, glucose oxidase, One enzyme, glucose oxidase, catalyzes the formation of gluconic catalyzes the formation of gluconic acid and hydrogen peroxide from acid and hydrogen peroxide from the oxidation of glucose. the oxidation of glucose.

• A second enzyme, peroxides A second enzyme, peroxides catalyzes the reaction of hydrogen catalyzes the reaction of hydrogen peroxide with potassium iodide peroxide with potassium iodide chromogen to oxidize the chromogen chromogen to oxidize the chromogen to colors ranging from green to to colors ranging from green to brown.brown.

KETONES IN URINE (ketonuria):KETONES IN URINE (ketonuria):•

fats--------carbon dioxide and water.fats--------carbon dioxide and water.• inadequate carbohydrate in the diet or a defect inadequate carbohydrate in the diet or a defect

in carbohydrate metabolism or absorption,in carbohydrate metabolism or absorption,• fatty acids------------metabolized. fatty acids------------metabolized. • When the fatty acid utilization is incomplete------- When the fatty acid utilization is incomplete-------

the intermediary products of fat metabolism the intermediary products of fat metabolism appear in the blood and the urine.appear in the blood and the urine.

• These products -------- acetone, These products -------- acetone, • diacetic acid (acetoacetic diacetic acid (acetoacetic

acid) betahydroxybutyric acid) betahydroxybutyric acid. acid.

• Diabetes mellitus, Diabetes mellitus, • Other Causes of Ketonuria:Other Causes of Ketonuria:

– FeverFever– AnorexiaAnorexia– Gastrointestinal disturbancesGastrointestinal disturbances– FastingFasting– StarvationStarvation– Severe vomiting Severe vomiting

• Rothera’s Test for Acetone and Rothera’s Test for Acetone and Acetoacetic Acid:Acetoacetic Acid:

• Procedure:Procedure:– Take 5ml of urine in a test tube and Take 5ml of urine in a test tube and

saturate it with ammonium sulphate. saturate it with ammonium sulphate. – Add 1 crystal of sodium nitroprusside.Run Add 1 crystal of sodium nitroprusside.Run

liquor ammonia carefully at the side of the liquor ammonia carefully at the side of the tube so as to form a layer on top of the tube so as to form a layer on top of the saturated urine.saturated urine.

Result: a permanganate calomel red Result: a permanganate calomel red (pink – purple) ring forms at the (pink – purple) ring forms at the junction of the two layers------positive.junction of the two layers------positive.

• Negative result shows no ring or a Negative result shows no ring or a brown ring.brown ring.

• Ferric Chloride Test for Diacetic Acid – Ferric Chloride Test for Diacetic Acid – Gerhardt’s TestGerhardt’s Test

• Harts Test for Beta-Hydroxybutyric Harts Test for Beta-Hydroxybutyric AcidAcid

• Colorimetric Reagent Strip Test Colorimetric Reagent Strip Test

• 3. Proteins in urine:3. Proteins in urine: Normal <30 mgms / 100 ml (30 – 50 Normal <30 mgms / 100 ml (30 – 50

mgms/24 hours)mgms/24 hours) Tests for Detection of ProteinsTests for Detection of Proteins• Semiquantitative precipitation Semiquantitative precipitation

tests: tests: 1.1.The heat and acetic acid method,The heat and acetic acid method,

2.sulfosalicyclic acid method and 2.sulfosalicyclic acid method and

3.the concentrated nitric acid protein 3.the concentrated nitric acid protein precipitation method precipitation. precipitation method precipitation.

• Negative – No turbidity or cloudiness.Negative – No turbidity or cloudiness.

• Trace - A faint precipitate visible against a Trace - A faint precipitate visible against a black background, equivalent to about 5 black background, equivalent to about 5 mg / dl protein.mg / dl protein.

• 1+ - Definite cloud without flocculation 1+ - Definite cloud without flocculation equivalent to 10 – 30 mg / dl.equivalent to 10 – 30 mg / dl.

• 2+ - Heavy and granular cloud without 2+ - Heavy and granular cloud without flocculation equivalent to 40 – 100 mg / dl.flocculation equivalent to 40 – 100 mg / dl.

• 3+ - dense cloud with marked 3+ - dense cloud with marked flocculation equivalent to 200 – 500 mg / dl flocculation equivalent to 200 – 500 mg / dl ..

• 4+ - cloudiness with precipitation 4+ - cloudiness with precipitation equivalent to 500 mg / dl or more.equivalent to 500 mg / dl or more.

• Heat and Acetic Acid Method Heat and Acetic Acid Method Procedure : Procedure : • Take a long test tube and fill ¾ the tube Take a long test tube and fill ¾ the tube

with clear urine.with clear urine.• Boil the upper portion over a flame .the Boil the upper portion over a flame .the

lower portion serves as the control.lower portion serves as the control.• If proteins, phosphates or carbonates are If proteins, phosphates or carbonates are

present in the urine a white cloud present in the urine a white cloud develops. develops.

• Add 1-3 drops of glacial acetic acid. Any Add 1-3 drops of glacial acetic acid. Any turbidity due to phosphate precipitation turbidity due to phosphate precipitation will clear or if it is due to carbonates they will clear or if it is due to carbonates they disappear with effervescence. disappear with effervescence.

• If it persists, it is due to albumin .If it persists, it is due to albumin .• precipitates due to mucin or precipitates due to mucin or

nucleoproteins will disappear on addition nucleoproteins will disappear on addition of 2 drops of nitric acid.of 2 drops of nitric acid.

• Sulphosalicylic acid methodSulphosalicylic acid method

• ProcedureProcedure

• Take 2 ml of clear urine in a test tube.Take 2 ml of clear urine in a test tube.

• Add an equal volume of 30 % Add an equal volume of 30 % sulphosalicylic acid.sulphosalicylic acid.

• Mix thoroughly, allow it to stand for 10 Mix thoroughly, allow it to stand for 10 minutes and estimate the amount of minutes and estimate the amount of turbidity.turbidity.

• Nitric acid method Nitric acid method • ProcedureProcedure • Take 2 – 3 ml of concentrated nitric acid in Take 2 – 3 ml of concentrated nitric acid in

a test tube.a test tube.• Carefully pour 5ml of clear urine down the Carefully pour 5ml of clear urine down the

inner side of the inclined test inner side of the inclined test • tube so that the urine forms a layer tube so that the urine forms a layer

over the nitric acid.over the nitric acid.• A ring of white precipitated protein will A ring of white precipitated protein will

form at the interface. Estimates the form at the interface. Estimates the amount of precipitate.amount of precipitate.

• Colorimetric reagent strip Colorimetric reagent strip testtest

• PRINCIPLE: the colorimetric reagent strip test is based PRINCIPLE: the colorimetric reagent strip test is based upon the ability of proteins to alter the color of some upon the ability of proteins to alter the color of some acid-base indicators without altering the pH. When an acid-base indicators without altering the pH. When an indicator such as indicator such as tetrabromphenol bluetetrabromphenol blue is buffered at pH is buffered at pH 3.it is yellow in solutions without protein. But in the 3.it is yellow in solutions without protein. But in the presence of protein the color will change to green and presence of protein the color will change to green and then to blue with increasing protein concentration. then to blue with increasing protein concentration.

• PROCEDURE: protein is determined by dipping the strip PROCEDURE: protein is determined by dipping the strip into well mixed un centrifuged urine and immediately into well mixed un centrifuged urine and immediately comparing the resultant color with the chart provided on comparing the resultant color with the chart provided on the reagent strip bottle.the reagent strip bottle.

• RESULTS: the results are reported as negative (yellow RESULTS: the results are reported as negative (yellow color), trace, 1+ to 4+.trace readings may detect 5 to 20 color), trace, 1+ to 4+.trace readings may detect 5 to 20 mg of protein/dl. Albumin reacts more strongly than the mg of protein/dl. Albumin reacts more strongly than the other proteins. Highly buffered alkaline urine and other proteins. Highly buffered alkaline urine and contamination of the urine specimen with quaternary contamination of the urine specimen with quaternary ammonium compounds or skin cleansers containing ammonium compounds or skin cleansers containing chlorohexidine may produce false results.chlorohexidine may produce false results.

Bence Jones ProteinuriaBence Jones Proteinuria: :

• Bence Jones protein isBence Jones protein is soluble at soluble at room and body temperatures. room and body temperatures.

• It precipitates upon heating between It precipitates upon heating between 45 C and 60 C 45 C and 60 C

• redissolves when the urine is further redissolves when the urine is further heated to the boiling point.heated to the boiling point.

• Reappear on coolingReappear on cooling

OCCULT BLOOD IN OCCULT BLOOD IN URINE:URINE: red blood cells red blood cells oror haemoglobinhaemoglobin

When hemolysis occurs When hemolysis occurs in circulationin circulation urineurine Normally an occasional red cell may be Normally an occasional red cell may be

found on microscopic examination of the found on microscopic examination of the urine sediment. In women during urine sediment. In women during menstruation, the urine may get menstruation, the urine may get contaminated with menstrual blood and contaminated with menstrual blood and hence examination of urine should not be hence examination of urine should not be done during that period.done during that period.

• Haematuria: Denotes the presence of Haematuria: Denotes the presence of red blood cells in urine. It is seen in red blood cells in urine. It is seen in various renal disorders, infectious or various renal disorders, infectious or neoplastic or trauma related to any part neoplastic or trauma related to any part of urinary tract.of urinary tract.

• Hemoglobinuria: is the presence of Hemoglobinuria: is the presence of blood pigments in the urine without the blood pigments in the urine without the presence of red blood cells. It is presence of red blood cells. It is associated with certain hemolytic associated with certain hemolytic anemia’s that cause hemolytic anemia, anemia’s that cause hemolytic anemia, transfusion reactions, malaria, and transfusion reactions, malaria, and paroxysmal nocturnal hemoglobinuriaparoxysmal nocturnal hemoglobinuria

• Tests for Detection of Blood:Tests for Detection of Blood:• 1. Microscopic Examination1. Microscopic Examination:: sediment------------sediment------------RBC’S/HPFRBC’S/HPF

• 2.2. Benzidine TestBenzidine Test PRINCIPLEPRINCIPLE Heme acts as a catalyst when hydrogen peroxide is Heme acts as a catalyst when hydrogen peroxide is

mixed with benzidine.mixed with benzidine. REAGENTSREAGENTS A: Saturated solution of benzidine in glacial acetic A: Saturated solution of benzidine in glacial acetic

acidacid B: Hydrogen peroxideB: Hydrogen peroxide PROCEDUREPROCEDURE Mix equal parts of A and B in a test tube and an Mix equal parts of A and B in a test tube and an

equal amount of the mixed reagent.equal amount of the mixed reagent.

RESULTSRESULTS• A blue color indicates the presence of A blue color indicates the presence of

hemoglobin.hemoglobin.

Colorimetric reagent strip Colorimetric reagent strip methodmethod

Principle: the reagent area is Principle: the reagent area is impregnated with impregnated with tetramethylbenzidine tetramethylbenzidine and buffered and buffered organic peroxide. This forms a green organic peroxide. This forms a green to dark blue compound when to dark blue compound when hemoglobin catalyzes the oxidation hemoglobin catalyzes the oxidation reaction of tetramethylbenzidine with reaction of tetramethylbenzidine with a peroxide.a peroxide.

BILE IN THE URINE:BILE IN THE URINE: The constituents The constituents 1.1. bilirubin (bile pigments),bilirubin (bile pigments),2.2. bile salts, bile salts, 3.3. urobilin and urobilinogen.urobilin and urobilinogen.

Bilirubin in appears IN JAUNDICEBilirubin in appears IN JAUNDICE

Bilirubin in Urine: bilirubin in the urine indicates the Bilirubin in Urine: bilirubin in the urine indicates the presence of hepatocellular disease or intra or extrahepatic presence of hepatocellular disease or intra or extrahepatic biliary obstruction biliary obstruction

in the reticuloendothelial cells in the reticuloendothelial cells breakdown of hemoglobin.breakdown of hemoglobin. linked to albumin----------linked to albumin---------- liver. This albumin-bound form, liver. This albumin-bound form,

which is also known as unconjugated bilirubin( indirect which is also known as unconjugated bilirubin( indirect bilirubin) insoluble in water and does not appear in the urine.bilirubin) insoluble in water and does not appear in the urine.

In the liver cells,--In the liver cells,--conjugated with glucuronic and sulfuric conjugated with glucuronic and sulfuric acids to form water soluble conjugated bilirubin (direct acids to form water soluble conjugated bilirubin (direct bilirubin). bilirubin).

It is secreted into the bile and then excreted into the It is secreted into the bile and then excreted into the intestinal tract through the bile duct. intestinal tract through the bile duct.

This conjugated bilirubin -------------This conjugated bilirubin -------------to urobilinogento urobilinogen. .

• certain liver diseases --certain liver diseases --unable to unable to conjugate all the bilirubin-----conjugate all the bilirubin-----an an increase in both conjugated and increase in both conjugated and unconjugated bilirubin --unconjugated bilirubin --bilirubinuria.bilirubinuria.

• obstructive biliary tract disease--obstructive biliary tract disease--bilirubinuria.bilirubinuria.

• hemolytic anemia’shemolytic anemia’s unconjugated unconjugated bilirubin--------bilirubin-------- urine free from urine free from bilirubin(Acholuric)bilirubin(Acholuric)

Tests for detection of bile Tests for detection of bile saltssalts::Hay Test Hay Test

Procedure: Procedure:

flowers of sulphur --sprinkled on the flowers of sulphur --sprinkled on the surface of the urine, surface of the urine,

Results:if bile salts are present they Results:if bile salts are present they sink to the bottom. sink to the bottom.

Otherwise they float on the surface. Otherwise they float on the surface.

This is due to the property of bile salts This is due to the property of bile salts to lower surface tensionto lower surface tension

Tests for detection of bile Tests for detection of bile pigments pigments

1.1. Foam Test Foam Test: : Shake urine in a test tube. If the foam on Shake urine in a test tube. If the foam on

top is yellow, bile pigments are present.top is yellow, bile pigments are present. 2. 2. Gmelins testGmelins test:: PROCEDURE:PROCEDURE: 1. Place ½ inch column of yellow nitric 1. Place ½ inch column of yellow nitric

acid in a test tube.acid in a test tube. 2. Overlay with equal amounts of urine 2. Overlay with equal amounts of urine

RESULT:RESULT: A play of colored rings, the most distinct A play of colored rings, the most distinct

being green indicates the presence of bile being green indicates the presence of bile pigments. pigments.

33. Fouchets Test. Fouchets Test::FOUCHETS REAGENTFOUCHETS REAGENT Trichloroacetic acid – 25 gmsTrichloroacetic acid – 25 gms Distilled water - 100 mlDistilled water - 100 ml 10% Ferric chloride solution – 10 ml.10% Ferric chloride solution – 10 ml.PROCEDUREPROCEDURE 1. Place 10 ml of acidified urine in a test 1. Place 10 ml of acidified urine in a test

tube.tube. 2. Add 2.5ml of 10 % barium chloride.2. Add 2.5ml of 10 % barium chloride. 3. Mix and filter.3. Mix and filter. 4. Unfold the filter paper and spread it on a 4. Unfold the filter paper and spread it on a

dry filter.dry filter. 5. Add 1 drop of Fouchets reagent to the 5. Add 1 drop of Fouchets reagent to the

residual precipitate.residual precipitate.RESULT: A green or blue color indicates the presence RESULT: A green or blue color indicates the presence

of bilirubin.of bilirubin.

Colorimetric Strip Reagent Colorimetric Strip Reagent TestTest• Principle: this test is base on the coupling Principle: this test is base on the coupling

of bilirubin with diazotized 2, 4- of bilirubin with diazotized 2, 4- • dichloroaniline in a strong acid dichloroaniline in a strong acid

medium to form a brown purple medium to form a brown purple azobilirubin azobilirubin

• Compound. The color ranges Compound. The color ranges through various shades of tan.through various shades of tan.

• Procedure:Procedure: the reagent strip is dipped the reagent strip is dipped into fresh, uncentrifuged urine tapped to into fresh, uncentrifuged urine tapped to

• remove excess urine and after remove excess urine and after 20 seconds, compared to the color chart 20 seconds, compared to the color chart

• on the reagent strip bottleon the reagent strip bottle• Result: the results are interpreted as Result: the results are interpreted as

negative, small (+), moderate (++), and negative, small (+), moderate (++), and large (+++) amounts of bilirubin.large (+++) amounts of bilirubin.

Tests for detection of Tests for detection of urobilinogenurobilinogen:: 1. Qualitative Ehrlich’s Test 1. Qualitative Ehrlich’s Test EHRLICH’S REAGENTEHRLICH’S REAGENT Paradimethylaminobenzaldehyde – 2 gmsParadimethylaminobenzaldehyde – 2 gms 20%5 HCL - 100 ml20%5 HCL - 100 ml PROCEDUREPROCEDURE a. Place 10 ml of urine in a test tube.a. Place 10 ml of urine in a test tube. b. Add 2.5 ml of barium chloride (to b. Add 2.5 ml of barium chloride (to

remove remove bilirubin). bilirubin). c. Mix well and filter.c. Mix well and filter. d. Add 0.5 ml of Ehrlich’s reagentd. Add 0.5 ml of Ehrlich’s reagent e. Allow it stand for 3 – 5 minutes.e. Allow it stand for 3 – 5 minutes.

Results:Results: pink colorpink color observable when viewed from the observable when viewed from the

top of the test tube. Against a white top of the test tube. Against a white background placed beneath the background placed beneath the bottom of the test tube. bottom of the test tube.

cherry red colorcherry red color Abnormally high amounts of This Abnormally high amounts of This

test must be done with fresh urine or test must be done with fresh urine or else urobilinogen is oxidized on else urobilinogen is oxidized on exposure to air urobilin. Excessively exposure to air urobilin. Excessively cold water should cold water should not be used in diluting the urine. not be used in diluting the urine.

MICROSCOPIC EXAMINATIONMICROSCOPIC EXAMINATION Qualitative technique: Qualitative technique:

the urine must be freshly voided the urine must be freshly voided

examined without excessive delay in order examined without excessive delay in order to prevent cellular deterioration. to prevent cellular deterioration.

Cellular debris from the urethral meatus and Cellular debris from the urethral meatus and secretions from the vagina may secretions from the vagina may contaminate the urine specimen. contaminate the urine specimen.

10-15 ml of urine ----10-15 ml of urine ----from freshly mixed from freshly mixed urine specimen and centrifuged at a urine specimen and centrifuged at a standard speed,standard speed,

usually 1500 to 2000 rpm for 5 minutes. usually 1500 to 2000 rpm for 5 minutes.

This is sufficient to bring to the bottom This is sufficient to bring to the bottom casts, pus cells, blood and crystals. For casts, pus cells, blood and crystals. For bacteria however a higher speed of 3,000 bacteria however a higher speed of 3,000 rpm is required. rpm is required.

the sediment resuspended in 1 ml.of the the sediment resuspended in 1 ml.of the same fluid.same fluid.

A drop of resuspended sediment is placed A drop of resuspended sediment is placed directly on a microscope slide and covered directly on a microscope slide and covered with a cover slip. with a cover slip.

. .

low power-low power-

Casts tend to congregate at the edges of Casts tend to congregate at the edges of the cover slip.the cover slip.

A minimum of A minimum of 10 – 15 high power fields10 – 15 high power fields should be scanned for this examination.should be scanned for this examination.

• Red blood cells, leucocytes,epithelial Red blood cells, leucocytes,epithelial cells---cells--- per high power field(/hpf); per high power field(/hpf);

• casts ---casts --- per low power fields(/lpf). per low power fields(/lpf).

Other elements such as Other elements such as bacteria, parasites, bacteria, parasites, crystals and spermatozoacrystals and spermatozoa are reported as are reported as wellwell

• NORMAL SEDIMENTNORMAL SEDIMENT

Normal sediment contains a limited Normal sediment contains a limited number of formed elements. it can number of formed elements. it can be divided into two classes.be divided into two classes.

• unorganized sedimentunorganized sediment

• Organized sedimentOrganized sediment

A. Unorganized sediment - these are the crystals of A. Unorganized sediment - these are the crystals of various substances present in the urine and they various substances present in the urine and they vary with the pH of the urine .crystals of normal vary with the pH of the urine .crystals of normal urine is formed as the specimen cools.urine is formed as the specimen cools.

1. 1. Crystals in acidic urine:Crystals in acidic urine:a.a. Uric acid and Urates;Uric acid and Urates; – – crystals are seen when the urine is allowed to crystals are seen when the urine is allowed to

stand for sometime and not seen in freshly stand for sometime and not seen in freshly passed urine. passed urine.

Amorphous uratesAmorphous urates appear as red granules and appear as red granules and are dissolved by heat and sodium hydroxide but are dissolved by heat and sodium hydroxide but not acetic acid. not acetic acid.

Uric acid crystalsUric acid crystals vary in shape and are yellow vary in shape and are yellow brown in colour and are not dissolved by heat, brown in colour and are not dissolved by heat, acetic acid or HCL but are soluble when heated acetic acid or HCL but are soluble when heated with sodium hydroxide. with sodium hydroxide.

disturbances of uric acid metabolism disturbances of uric acid metabolism fevers where the urine is concentrated.fevers where the urine is concentrated.

Acid uric crystals

B.B.Calcium Oxalate:Calcium Oxalate: They are commonly found in diets rich in They are commonly found in diets rich in

tomatoestomatoes, , spinach spinach etc. they are typically etc. they are typically envelope shapedenvelope shaped crystals but occasionally crystals but occasionally appear dumb- bell shaped .they are appear dumb- bell shaped .they are insoluble in strong Hcl. insoluble in strong Hcl.

c. Cystine Crystalsc. Cystine Crystals: : highly refractile, highly refractile, hexagonal plateshexagonal plates and are and are

soluble in Hcl but insoluble in acetic acid. soluble in Hcl but insoluble in acetic acid. They are seen They are seen cystinosis cystinosis which is an which is an inborn error of metabolism in which inborn error of metabolism in which cystine crystals are found in the urine, cystine crystals are found in the urine, reticuloendothelial system and eyes. reticuloendothelial system and eyes.

d.d.LeucineLeucine:: slightly yellowslightly yellow, oily looking , oily looking spheresspheres with with

radial and concentric striations .they are radial and concentric striations .they are not soluble in Hcl or ether .they are found not soluble in Hcl or ether .they are found in in liver disordersliver disorders..

Calcium OxalateCalcium Oxalate

Cystine CrystalsCystine Crystals

Leucine CrystalsLeucine Crystals

• e.e.TyrosineTyrosine:– :– fine needlesfine needles arranged in concentric sheaves, constriction at the arranged in concentric sheaves, constriction at the

middle. middle. liver disorders.liver disorders.

• f.f.Sulpha crystalsSulpha crystals :- :- patients taking sulphonamides. patients taking sulphonamides.

2. Crystals in alkaline urine:2. Crystals in alkaline urine:

• a. Ammonium Magnesium Phosphatesa. Ammonium Magnesium Phosphates: : ( triple phosphate) ( triple phosphate) coffin lid, feathery or leaf like formscoffin lid, feathery or leaf like forms. . In freshly passed urine they indicate stones in the bladder or In freshly passed urine they indicate stones in the bladder or

kidney.kidney. Phosphates may occur as amorphous deposits in alkaline urine Phosphates may occur as amorphous deposits in alkaline urine

and are dissolved in acetic acid.and are dissolved in acetic acid.• b. b. Dicalcium PhosphatesDicalcium Phosphates: hey are also seen in slightly acid or : hey are also seen in slightly acid or

neutral urine. They are neutral urine. They are colorless prismscolorless prisms arranged in arranged in stars and stars and rosettesrosettes

They are soluble in acetic acid.They are soluble in acetic acid. c. c. Calcium carbonateCalcium carbonate: amorphous granules or colorless spheres : amorphous granules or colorless spheres

and dumb-bells which are soluble in acetic acid with gas and dumb-bells which are soluble in acetic acid with gas formation.formation.

Tyrosine crystalsTyrosine crystals

Ammonium Magnesium Ammonium Magnesium PhosphatesPhosphates

Ammonium Biurate CrystalsAmmonium Biurate Crystals

Calcium PhosphateCalcium Phosphate

Crystals in acidCrystals in acid urineurine Amorphous urates-Amorphous urates-Brick – Red-Brick – Red-

GranulesGranules Uric AcidUric Acid - -Yellow – Brown--Yellow – Brown--

Polymorphous – Whetstones,Rosettes Polymorphous – Whetstones,Rosettes of prisms,Rhombohedral prisms, of prisms,Rhombohedral prisms, hexagonal platehexagonal plate

Sodium urateSodium urateColorless to YellowColorless to YellowFan Fan of slender prisms of slender prisms

Cystine (rare)Cystine (rare)1. Colorless2.Highly 1. Colorless2.Highly refractilerefractileFlat hexagonal plates with Flat hexagonal plates with well – defined edges singly or in well – defined edges singly or in clustersclusters

Cholestrol (rare)Cholestrol (rare) - -ColorlessBroken window ColorlessBroken window panes with notched corners ,Flat plates panes with notched corners ,Flat plates

Leucine (rare)-Leucine (rare)-1. Yellow or Brown2.Highly 1. Yellow or Brown2.Highly refractile-refractile-Spheroids with striations pure Spheroids with striations pure form hexagonal form hexagonal

Tyrosine (rare)Tyrosine (rare) Colorless or YellowFine Colorless or YellowFine silky needles in sheaves or rosettes silky needles in sheaves or rosettes

BilirubinBilirubin--Reddish Brown--Reddish Brown--Cubes, Cubes, Rhombic plates, Amorphous needles Rhombic plates, Amorphous needles

Cholesterol CrystalsCholesterol Crystals

Acid, Neutral or Slightly Acid, Neutral or Slightly Alkaline UrineAlkaline Urine

Calcium oxalate-Calcium oxalate---ColorlessColorless Octahedral Dumbbells,Often small Octahedral Dumbbells,Often small

Hippuric acidHippuric acid Colorless- Colorless- Rhombic Rhombic plates Four sided prismsplates Four sided prisms

Alkaline UrineAlkaline Urine Calcium Carbonate-Calcium Carbonate-Colorless --Colorless --

Needles,Spheres,Dumbbells Needles,Spheres,Dumbbells Ammonium biurate-Ammonium biurate-Yellow Opaque Yellow Opaque

BrownThorn BrownThorn apple,Spheres,Dumbbells,Sheaves of apple,Spheres,Dumbbells,Sheaves of needles needles

Calcium Phosphate-Calcium Phosphate-Colorless-Colorless- Prisms, Plates, Needles - - - -Prisms, Plates, Needles - - - -

A- represents the A- represents the residueresidue of normal human urine, as seen under the microscope. of normal human urine, as seen under the microscope.

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

Nitrate of urea is represented in division C. Nitrate of urea is represented in division C.

A and B--highly magnified urinary deposits, which indicate A and B--highly magnified urinary deposits, which indicate different degrees of impairment of the digestive functions different degrees of impairment of the digestive functions are represented. are represented.

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

indicative of disease of the mucous lining of the urinary organs.indicative of disease of the mucous lining of the urinary organs.

epithelial cellsepithelial cells

• In division A are represented the mixed urates as they In division A are represented the mixed urates as they appear during idiopathic fevers, as intermittent, remittent, appear during idiopathic fevers, as intermittent, remittent, etc. When appearing as seen in division B, a less violent etc. When appearing as seen in division B, a less violent affection of the same character is indicated. Division C affection of the same character is indicated. Division C represents urate of ammonia, occasionally observed when represents urate of ammonia, occasionally observed when there is a tendency towards albuminuria, In division D there is a tendency towards albuminuria, In division D which is present in the urine of persons suffering from gout. which is present in the urine of persons suffering from gout. The crystals shown in division E consist of the same salt. The crystals shown in division E consist of the same salt.

Uric acidUric acid

varieties of cancer. varieties of cancer.

• Organized SedimentOrganized Sediment: the components of : the components of organized sediment include casts, red blood cells, organized sediment include casts, red blood cells, white blood cells, epithelial cells, bacteria, yeast, white blood cells, epithelial cells, bacteria, yeast, parasites, spermatozoa and artifacts.parasites, spermatozoa and artifacts.

• a. a. Casts:Casts: • Casts are formed in the tubules and is composed Casts are formed in the tubules and is composed

of of proteinaceous materialproteinaceous material. They are washed out . They are washed out by the glomerular secretion into the collecting by the glomerular secretion into the collecting tubules and the bladder. They are tubules and the bladder. They are cylindrical in cylindrical in shapeshape with with round or broken endsround or broken ends with uniform with uniform diameter but varying in length. They require diameter but varying in length. They require acidic conditions, high salt concentration, acidic conditions, high salt concentration, reduced urine flow and protein to be formed. reduced urine flow and protein to be formed. Practically all casts have a Practically all casts have a hyaline matrixhyaline matrix, which , which may or may not contain inclusions such as may or may not contain inclusions such as desquamated cells.desquamated cells.

• The casts are The casts are named according to the matrix of named according to the matrix of the inclusionsthe inclusions contained in them e.g. red blood contained in them e.g. red blood cell casts.cell casts.

1. 1. Hyaline casts:Hyaline casts: are colorless, are colorless, semi-transparent andsemi-transparent and occasionally refractile cylinders and are soluble in occasionally refractile cylinders and are soluble in

acetic acid. They are seen when there is damage to acetic acid. They are seen when there is damage to the glomerular capillary membrane, permitting the glomerular capillary membrane, permitting leakage of proteins through the glomerular filtrate. leakage of proteins through the glomerular filtrate. They are seen in They are seen in fever, orthostatic proteinuria, and fever, orthostatic proteinuria, and emotional stress or strenuous exercise.emotional stress or strenuous exercise.

2. 2. Granular castsGranular casts: are casts containing large or fine : are casts containing large or fine granules embedded in coagulated protein. They are granules embedded in coagulated protein. They are not found in normal urine and their presences not found in normal urine and their presences indicate indicate pyelonephritis.pyelonephritis. They are also seen in They are also seen in chronic lead poisoning.chronic lead poisoning.

3. 3. Epithelial castsEpithelial casts: are formed of fused desquamated : are formed of fused desquamated tubular cells. They are coagulated protein in which tubular cells. They are coagulated protein in which are embedded desquamated epithelial cells from are embedded desquamated epithelial cells from the renal tubules .they are seen in diseases where the renal tubules .they are seen in diseases where there is damage to the tubular epithelium as in there is damage to the tubular epithelium as in nephrosis, eclampsia, amyloidosis and heavy metal nephrosis, eclampsia, amyloidosis and heavy metal poisoningpoisoning. .

Hyaline castsHyaline casts

Granular castsGranular casts

4. 4. Red Blood Cell CastsRed Blood Cell Casts::

are casts with red blood cells are casts with red blood cells embedded in the coagulated protein embedded in the coagulated protein in the tubule. Their presences indicate in the tubule. Their presences indicate acute inflammationacute inflammation or vascular or vascular disorder in the glomerulus causing disorder in the glomerulus causing hematuriahematuria. They are seen in . They are seen in pathological conditions such as pathological conditions such as acute acute glomerulonephritisglomerulonephritis, , renal infarctionrenal infarction and collagen vascular disorder. and collagen vascular disorder.

5. 5. White Blood Cell Casts (Pus cell)White Blood Cell Casts (Pus cell)

6. 6. Fatty CastsFatty Casts

7. 7. Waxy CastsWaxy Casts

Red Blood Cell CastsRed Blood Cell Casts

White Blood Cell CastsWhite Blood Cell Casts

Waxy CastsWaxy Casts

• CELLS:CELLS:• a. a. Red blood cells:Red blood cells: Normally 1-2 red blood Normally 1-2 red blood

cell are found per high power fieldcell are found per high power field• .they appear pale , light refractive, biconcave .they appear pale , light refractive, biconcave

discs when viewed under high power discs when viewed under high power magnification .they have no nuclei. Red blood magnification .they have no nuclei. Red blood cells in fresh, unstained sediment appear pale cells in fresh, unstained sediment appear pale in color; in urine that is not fresh, they are in color; in urine that is not fresh, they are pale or colorless shadow cells .in concentrated pale or colorless shadow cells .in concentrated urine, they may be small and crenated; and in urine, they may be small and crenated; and in dilute urine, they are large and swollen and dilute urine, they are large and swollen and sometimes rupture to produce ghost cells.sometimes rupture to produce ghost cells.

• b. b. White cellsWhite cells• c. c. Epithelial CellsEpithelial Cells: Normally a few epithelial : Normally a few epithelial

cells occur in the urine .A marked increasecells occur in the urine .A marked increase• is these cells in the urine is seen is these cells in the urine is seen

destruction of the tissues in the urinary tract.destruction of the tissues in the urinary tract.

Red blood cellsRed blood cells

Red blood cellsRed blood cells

White cellsWhite cells

White cellsWhite cells

Squamous Squamous Epithelial Epithelial CellsCells

Transitional Epithelial Transitional Epithelial CellsCells

Renal Tubular Epithelial Renal Tubular Epithelial CellsCells

• Quantitative Evaluation of the Quantitative Evaluation of the urine sediment – Addis counturine sediment – Addis count

• The Addis countThe Addis count is a quantitative is a quantitative measurement of the excretion of red measurement of the excretion of red cells, leucocytes and casts in the cells, leucocytes and casts in the urine during a 12 hour period.urine during a 12 hour period.

• e. Bacteria:e. Bacteria:

Bacteruria is considered significant Bacteruria is considered significant when there is the presence of when there is the presence of 100,000100,000 or more bacteria per ml of or more bacteria per ml of urine specimen.urine specimen.

DETECTION OF BACTERIADETECTION OF BACTERIA• Microscopic Examination:Microscopic Examination: sediment -sediment - > >20 or more bacteria per high 20 or more bacteria per high

power fieldpower field may indicate a urinary tract may indicate a urinary tract infectioninfection

Reagent stripsReagent strips:: PRINCIPLE:PRINCIPLE: This test depends upon the conversion of This test depends upon the conversion of

nitrate to nitritenitrate to nitrite by the action of gram by the action of gram negative bacteria in urine. At the acid pH of negative bacteria in urine. At the acid pH of the reagent area, nitrite in the urine reacts the reagent area, nitrite in the urine reacts with p-arsanilic acid to form diazonium with p-arsanilic acid to form diazonium compound. This compound in turn couples compound. This compound in turn couples with 1, 2, 3, 4-tetra hydrobenzoquinolin-3-ol to with 1, 2, 3, 4-tetra hydrobenzoquinolin-3-ol to produce pink color.produce pink color.

• Procedure:Procedure: The strip is dipped in the urine The strip is dipped in the urine specimen for 5 seconds.specimen for 5 seconds.

Results:Results: uniform pink color --uniform pink color --positive result positive result

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

A negative result should never be interpreted as indicating A negative result should never be interpreted as indicating absence of bacteruria.absence of bacteruria.

There are several reasons for this.There are several reasons for this.– First morning urine or urine that has remained in the First morning urine or urine that has remained in the

bladder for several hours is more likely to yield a bladder for several hours is more likely to yield a positive nitrite test result in the presence of significant positive nitrite test result in the presence of significant bacteruria than bacteruria than a random urine samplea random urine sample that may have that may have been in the bladder for short time. in the latter type of been in the bladder for short time. in the latter type of specimen there may have been insufficient time(less specimen there may have been insufficient time(less than 4 hours) for conversion of nitrate to nitrite to have than 4 hours) for conversion of nitrate to nitrite to have occurred.occurred.

– When dietary nitrate is absentWhen dietary nitrate is absent, even if organisms , even if organisms containing reductase are present and bladder incubation containing reductase are present and bladder incubation is ample.is ample.

– Ascorbic acidAscorbic acid concentrations of more than 25 mg / dl or concentrations of more than 25 mg / dl or greater may cause false negative results with specimens greater may cause false negative results with specimens containing small amounts of nitrate.containing small amounts of nitrate.