Urine Presentation

URINE ANALYSISURINE ANALYSIS

DR.P.G.KONAPURDR.P.G.KONAPUR PROFESSOR OF PATHOLOGYPROFESSOR OF PATHOLOGY V.M.K.V.MEDICAL COLLEGEV.M.K.V.MEDICAL COLLEGE SALEMSALEM

11www.similima.comwww.similima.com

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 of Information--- about the functioning of

the kidney the kidney • diagnosis ---of urinary tract diseases diagnosis ---of urinary tract diseases • diagnosis -----of certain metabolic and diagnosis -----of certain metabolic and

systemic diseases. systemic diseases.


• Urine is formed in the kidney. Urine is formed in the kidney. • The functional unit of kidney is called nephrons,The functional unit of kidney is called 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 and followed by absorption of most of the water and

some of the solutes. 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 COMPOSITION 3 NORMAL CONSTITUENTS AND COMPOSITION OF URINEOF 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 mouthed, clean Containers for collection of urine should be wide mouthed, clean

and dryand 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, electrolytes, and quantitative estimation of proteins, sugars, electrolytes, and

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

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

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


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

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

swab. 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 etc .if concentrated samples and casts, crystals etc .if present present


• Terminal urine sample patient voids the last portion of Terminal urine sample patient voids the last portion of urine in an open container.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 the in place for 1- 3 hours, depending on the examination requested. 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 on Toluene – add a few drops till it forms a thin layer on the surface of urine.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. 99www.similima.comwww.similima.com

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 day. is total suppression of urine <100 ml per day.• Appearance – color & turbidityAppearance – color & turbidity• 1. Color1. Color - normal ---- amber yellow ( to the presence - normal ---- amber yellow ( to the presence

of urobilin,uroerythrin,and urochromes)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 & uric acid 4.Cloudy--------Phosphates & carbonates, Urates & uric acid

Pus cells 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),MelaninMethemoglobin,,Homogenesticacid(alkaptonuria),Melanin 7. orange--------Bile pigments,Drugs – pyridium,rifampicin7. orange--------Bile pigments,Drugs – pyridium,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 or amorphous phosphate and carbonates – alkaline 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 strong forming ammonia which gives a strong ammonical smell.. 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 which Normal kidneys are capable of producing urine the pH of which

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

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

• PROCEDUREPROCEDURE

• Dip the litumus paper strips in the urine, remove and read the color Dip the litumus paper strips in the urine, remove and read the color change immediately.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 which nitrazine paper method :The Ph---- nitrazine paper which

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


SPECIFIC GRAVITYSPECIFIC GRAVITY

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

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

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

ESTIMATION OF SPECIFIC GRAVITY BY ESTIMATION OF SPECIFIC GRAVITY BY URINOMETERURINOMETER

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

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


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

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

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

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

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

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

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

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


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

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

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

• substract 0.001 for each 3 C, the temperature below the substract 0.001 for each 3 C, the temperature below the standard temperature.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 = 1.0051.001 = 1.005• Correction is also recommended when glucose or protein Correction is also recommended when glucose or protein

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


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

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

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

• This test is not specific for sugars and is affected by most of the This test is not specific for sugars and is affected by most of the reducing substances. 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 ml of Dissolve crystalline copper sulphate in 100 ml of distilled water. Dissolve sodium carbonate and distilled water. Dissolve sodium carbonate and sodium citrate in 700 cc. of distilled water. sodium citrate in 700 cc. of distilled water. Slowly add the latter to the former solution with Slowly add the latter to the former solution with constant stirring. When complete, make up the constant stirring. When complete, make up the volume to 1000ml with distilled water.volume to 1000ml 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 slightly Trace - solution appears pale green to slightly

cloudy.cloudy. 1+ - Definite cloudy green (0.5% sugar)1+ - Definite cloudy green (0.5% sugar) 2+ - Yellow to orange precipitate, supernatant 2+ - Yellow to orange precipitate, supernatant

fluid pale blue (1% sugar)fluid pale blue (1% sugar) 3+ - Orange to red precipitate, supernatant fluid 3+ - Orange to red precipitate, supernatant fluid

pale blue (1.5% sugars) pale blue (1.5% sugars) 4+ - Brick red precipitate, supernatant fluid 4+ - Brick red precipitate, supernatant fluid

decolorizes (2 % sugar) decolorizes (2 % sugar) 2222www.similima.comwww.similima.com

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

• A second enzyme, peroxides catalyzes A second enzyme, peroxides catalyzes the reaction of hydrogen peroxide with the reaction of hydrogen peroxide with potassium iodide chromogen to oxidize potassium iodide chromogen to oxidize the chromogen to colors ranging from the chromogen to colors ranging from green to brown.green to 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 in inadequate carbohydrate in the diet or a defect in

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

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

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

betahydroxybutyric acid. betahydroxybutyric 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 saturate Take 5ml of urine in a test tube and saturate

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

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

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

• Harts Test for Beta-Hydroxybutyric AcidHarts Test for Beta-Hydroxybutyric Acid• 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 tests: Semiquantitative precipitation 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 mg / dl black background, equivalent to about 5 mg / dl protein.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 flocculation 3+ - dense cloud with marked flocculation equivalent to 200 – 500 mg / dl .equivalent to 200 – 500 mg / dl .

• 4+ - cloudiness with precipitation equivalent 4+ - cloudiness with precipitation equivalent to 500 mg / dl or more.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 with Take a long test tube and fill ¾ the tube with

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

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

present in the urine a white cloud develops. present in the urine a white cloud develops. • Add 1-3 drops of glacial acetic acid. Any Add 1-3 drops of glacial acetic acid. Any

turbidity due to phosphate precipitation will turbidity due to phosphate precipitation will clear or if it is due to carbonates they 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 nucleoproteins will precipitates due to mucin or nucleoproteins will

disappear on addition of 2 drops of nitric acid.disappear on addition 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 a Take 2 – 3 ml of concentrated nitric acid in a

test tube.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 over tube so that the urine forms a layer over

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

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


• Colorimetric reagent strip testColorimetric reagent strip test

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

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

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


Bence Jones ProteinuriaBence Jones Proteinuria: :

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

• It precipitates upon heating between 45 C It precipitates upon heating between 45 C and 60 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 URINE:OCCULT BLOOD IN 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 found Normally an occasional red cell may be found

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


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

• Hemoglobinuria: is the presence of blood Hemoglobinuria: is the presence of blood pigments in the urine without the presence of pigments in the urine without the presence of red blood cells. It is associated with certain red blood cells. It is associated with certain hemolytic anemia’s that cause hemolytic hemolytic anemia’s that cause hemolytic anemia, transfusion reactions, malaria, and anemia, 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 acidA: Saturated solution of benzidine in glacial acetic acid 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 hemoglobin.A blue color indicates the presence of hemoglobin.


Colorimetric reagent strip Colorimetric reagent strip methodmethod

Principle: the reagent area is impregnated Principle: the reagent area is impregnated with with tetramethylbenzidine tetramethylbenzidine and buffered and buffered organic peroxide. This forms a green to organic peroxide. This forms a green to dark blue compound when hemoglobin dark blue compound when hemoglobin catalyzes the oxidation reaction of catalyzes the oxidation reaction of tetramethylbenzidine with a peroxide.tetramethylbenzidine with 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 presence of Bilirubin in Urine: bilirubin in the urine indicates the presence of hepatocellular disease or intra or extrahepatic biliary obstruction hepatocellular disease or intra or extrahepatic 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 bilirubin) which is also known as unconjugated bilirubin( indirect bilirubin) insoluble in water and does not appear in the urine.insoluble in water and does not appear in the urine.

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

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

This conjugated bilirubin -------------This conjugated bilirubin -------------to urobilinogento urobilinogen. . 4141www.similima.comwww.similima.com

• certain liver diseases --certain liver diseases --unable to unable to conjugate all the bilirubin-----conjugate all the bilirubin-----an increase an increase in both conjugated and unconjugated in both conjugated and unconjugated bilirubin --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 saltsTests for detection of bile salts::Hay Test Hay Test Procedure: Procedure: flowers of sulphur --sprinkled on the surface flowers of sulphur --sprinkled on the surface

of the urine, of the urine, Results:if bile salts are present they sink to Results:if bile salts are present they sink to

the bottom. the bottom. Otherwise they float on the surface. Otherwise they float on the surface. This is due to the property of bile salts to This is due to the property of bile salts to

lower surface tensionlower surface tension


Tests for detection of bile pigments Tests for detection of bile 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 acid 1. Place ½ inch column of yellow nitric acid

in a test tube.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 being A play of colored rings, the most distinct being

green indicates the presence of bile pigments. green indicates the presence of bile pigments. 4444www.similima.comwww.similima.com

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 tube.1. Place 10 ml of acidified urine in a test 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 dry 4. Unfold the filter paper and spread it on a dry

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

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

bilirubin.bilirubin.


Colorimetric Strip Reagent TestColorimetric Strip Reagent Test• Principle: this test is base on the coupling of Principle: this test is base on the coupling of

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

medium to form a brown purple azobilirubin medium to form a brown purple 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 into the reagent strip is dipped into

fresh, uncentrifuged urine tapped to fresh, uncentrifuged urine tapped to • remove excess urine and after 20 remove excess urine and after 20

seconds, compared to the color chart seconds, compared to the color chart • on the reagent strip bottleon the reagent strip bottle• Result: the results are interpreted as negative, Result: the results are interpreted as negative,

small (+), moderate (++), and large (+++) small (+), moderate (++), and large (+++) amounts of bilirubin.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 remove b. Add 2.5 ml of barium chloride (to 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 top of observable when viewed from the top of

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

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

must be done with fresh urine or else must be done with fresh urine or else urobilinogen is oxidized on exposure to air urobilinogen is oxidized on exposure to air urobilin. Excessively cold water should urobilin. Excessively cold water should not be used in diluting not be used in diluting the urine. the urine. 4848www.similima.comwww.similima.com

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

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


10-15 ml of urine ----10-15 ml of urine ----from freshly mixed urine from freshly mixed urine specimen and centrifuged at a standard speed,specimen and centrifuged at a 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 casts, pus This is sufficient to bring to the bottom casts, pus cells, blood and crystals. For bacteria however a cells, blood and crystals. For bacteria however a higher speed of 3,000 rpm is required. higher speed of 3,000 rpm is required.

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

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

. .


low power-low power- Casts tend to congregate at the edges of the Casts tend to congregate at the edges of the

cover slip.cover slip. A minimum of A minimum of 10 – 15 high power fields10 – 15 high power fields should should

be scanned for this examination.be scanned for this examination.

• Red blood cells, leucocytes,epithelial cells---Red blood cells, leucocytes,epithelial 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 well are reported as well


• NORMAL SEDIMENTNORMAL SEDIMENT Normal sediment contains a limited number Normal sediment contains a limited number

of formed elements. it can be divided into of formed elements. it can be divided into two classes.two classes.

• unorganized sedimentunorganized sediment

• Organized sedimentOrganized sediment


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

sometime and sometime and are not seen in freshly passed urine. are not seen in freshly passed urine. Amorphous uratesAmorphous urates appear as red granules and are appear as red granules and are

dissolved by heat and sodium hydroxide but not acetic dissolved by heat and sodium hydroxide but not acetic acid. acid.

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

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


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

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

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


• 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 formation.and dumb-bells which are soluble in acetic acid with gas formation.5555www.similima.comwww.similima.com

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 of Polymorphous – Whetstones,Rosettes of prisms,Rhombohedral prisms, hexagonal prisms,Rhombohedral prisms, hexagonal plateplate

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

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


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 form Spheroids with striations pure form hexagonal hexagonal

Tyrosine (rare)Tyrosine (rare) Colorless or YellowFine silky Colorless or YellowFine silky

needles in sheaves or rosettes needles in sheaves or rosettes

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


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

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

Hippuric acidHippuric acid Colorless- Colorless- Rhombic plates Rhombic plates Four sided prismsFour 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, Prisms, Plates, Needles - - - -Plates, Needles - - - -


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

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

Nitrate of urea is represented in division C. Nitrate of urea is represented in division C. 6060www.similima.comwww.similima.com

A and B--highly magnified urinary deposits, which indicate different A and B--highly magnified urinary deposits, which indicate different degrees of impairment of the digestive functions are represented. degrees of impairment of the digestive functions 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 shown globules are alone represented. In the division marked C are shown blood corpuscles as they are arranged in blood drawn from a vein or blood corpuscles as they are arranged in blood drawn from a vein or artery. D represents the same separated, as they always are when artery. D represents the same separated, as they always are when present in the urine. In E highly magnified oil globules are represented. present in the urine. In E highly magnified oil globules are represented. If present in the urine, they indicate disease of the kidneys. In the If present in the urine, they indicate disease of the kidneys. In the division marked F are represented epithelial cells, the presence of division marked F are represented epithelial cells, the presence of which in large numbers is indicative of disease of the mucous lining of which in large numbers is indicative of disease of the mucous lining of

the urinary organs.the urinary organs.

epithelial cellsepithelial cells


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

Uric acidUric acid


varieties of cancer. varieties of cancer.


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

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

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

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


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

acid. They are seen when there is damage to the acid. They are seen when there is damage to the glomerular capillary membrane, permitting leakage of glomerular capillary membrane, permitting leakage of proteins through the glomerular filtrate. They are seen proteins through the glomerular filtrate. They are seen in in fever, orthostatic proteinuria, and emotional stress or fever, orthostatic proteinuria, and emotional stress or strenuous exercise.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 not granules embedded in coagulated protein. They are not found in normal urine and their presences indicate found in normal urine and their presences indicate pyelonephritis.pyelonephritis. They are also seen in chronic lead They are also seen in chronic lead poisoning.poisoning.

3. 3. Epithelial castsEpithelial casts: are formed of fused desquamated : are formed of fused desquamated tubular cells. They are coagulated protein in which are tubular cells. They are coagulated protein in which are embedded desquamated epithelial cells from the renal embedded desquamated epithelial cells from the renal tubules .they are seen in diseases where there is tubules .they are seen in diseases where there is damage to the tubular epithelium as in damage to the tubular epithelium as in nephrosis, nephrosis, eclampsia, amyloidosis and heavy metal poisoningeclampsia, amyloidosis and heavy metal poisoning. . 6969www.similima.comwww.similima.com

4. 4. Red Blood Cell CastsRed Blood Cell Casts:: are casts with red blood cells embedded are casts with red blood cells embedded

in the coagulated protein in the tubule. in the coagulated protein in the tubule. Their presences indicate Their presences indicate acute acute inflammationinflammation or vascular disorder in the or vascular disorder in the glomerulus causing glomerulus causing hematuriahematuria. They are . They are seen in pathological conditions such as seen in pathological conditions such as acute glomerulonephritisacute glomerulonephritis, , 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 Casts7. 7. Waxy CastsWaxy Casts


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

are found per high power fieldare 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 in cells in fresh, unstained sediment appear pale in color; in urine that is not fresh, they are pale or color; in urine that is not fresh, they are pale or colorless shadow cells .in concentrated urine, colorless shadow cells .in concentrated urine, they may be small and crenated; and in dilute they may be small and crenated; and in dilute urine, they are large and swollen and sometimes urine, they are large and swollen and sometimes rupture to produce ghost cells.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 destruction is these cells in the urine is seen destruction

of the tissues in the urinary tract.of the tissues in the urinary tract.7171www.similima.comwww.similima.com

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

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

• e. Bacteria:e. Bacteria: Bacteruria is considered significant when Bacteruria is considered significant when

there is the presence of there is the presence of 100,000100,000 or more or more bacteria per ml of urine specimen.bacteria per ml of 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 infection may indicate a urinary tract infection Reagent stripsReagent strips:: PRINCIPLE:PRINCIPLE: This test depends upon the conversion of This test depends upon the conversion of nitrate nitrate

to nitriteto nitrite by the action of gram negative bacteria by the action of gram negative bacteria in urine. At the acid pH of the reagent area, in urine. At the acid pH of the reagent area, nitrite in the urine reacts with p-arsanilic acid to nitrite in the urine reacts with p-arsanilic acid to form diazonium compound. This compound in form diazonium compound. This compound in turn couples with 1, 2, 3, 4-tetra turn couples with 1, 2, 3, 4-tetra hydrobenzoquinolin-3-ol to produce pink color.hydrobenzoquinolin-3-ol to 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. 7373www.similima.comwww.similima.com

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

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

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

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

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


Urine Presentation

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