Culturing of Urine Skills in Microbiology

8/7/2019 Culturing of Urine Skills in Microbiology

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Dr.T.V.Rao.MDDr.T.V.Rao.MDDr.T.V.Rao.MDDr.T.V.Rao.MD

CULTURING OF URINESKILL BASED LEARNING

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OVER VIEW OF URINARY TRACTOVER VIEW OF URINARY TRACTOVER VIEW OF URINARY TRACTOVER VIEW OF URINARY TRACTINFECTIONSINFECTIONSINFECTIONSINFECTIONS

Most common infectious diseaseMost Numerous specimens are received in the Laboratory30 to 40 % of specimens received in Microbiologylaboratories are Urine specimens, to Identify the Infection.Diagnostic information is important for the clinician.Appropriate clinical information gives many clues for better diagnostic evaluations .

Specimen collection is the primary objective in getting anideal sample.

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an infection of one or

more structures in theurinary system. Most UTIsare caused by gram-negative bacteria, mostcommonly Escherichiacoli or species ofKlebsiella, Proteus,Pseudomonas, orEnterobacter, althoughother strains, such asStaphlyococcus andSerratia, are emerging

WHAT IS URINARY TRACT INFECTIONWHAT IS URINARY TRACT INFECTIONWHAT IS URINARY TRACT INFECTIONWHAT IS URINARY TRACT INFECTION

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The condition is more

common in women thanin men. UTI may beasymptomatic but isusually characterized byurinary frequency, burningpain with voiding, and, ifthe infection is severe,

visible blood and pus inthe urine. Fever and backpain often acco mpany kidneyinfections

URINARY TRACT INFECTIONS ARE MORECOMMON IN WOMEN

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COMMON UROPATHOGENSCOMMON UROPATHOGENSCOMMON UROPATHOGENSCOMMON UROPATHOGENS

Escherichia coli Escherichia coli Escherichia coli Escherichia coli

OtherOtherOtherOther EnterobacteriaceaeEnterobacteriaceaeEnterobacteriaceaeEnterobacteriaceae ((((KlebsiellaKlebsiellaKlebsiellaKlebsiella ,,,, Enterobacter Enterobacter Enterobacter Enterobacter ,,,,ProteusProteusProteusProteus ,,,, Citrobacter Citrobacter Citrobacter Citrobacter ))))Pseudomonas aeruginosaPseudomonas aeruginosaPseudomonas aeruginosaPseudomonas aeruginosaEnterococcusEnterococcusEnterococcusEnterococcusStaphylococcus saprophyticusStaphylococcus saprophyticusStaphylococcus saprophyticusStaphylococcus saprophyticusStaphylococcus aureusStaphylococcus aureusStaphylococcus aureusStaphylococcus aureusStreptococcus agalactiaeStreptococcus agalactiaeStreptococcus agalactiaeStreptococcus agalactiae (group B)(group B)(group B)(group B)

CandidaCandidaCandidaCandidaAssociated with staphylococcemiaAssociated with staphylococcemiaAssociated with staphylococcemiaAssociated with staphylococcemiaDenotes vaginal colonization in pregnant womenDenotes vaginal colonization in pregnant womenDenotes vaginal colonization in pregnant womenDenotes vaginal colonization in pregnant women

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CorynebacteriumCorynebacteriumCorynebacteriumCorynebacteriumurealyticumurealyticumurealyticumurealyticum 1111HaemophilusHaemophilusHaemophilusHaemophilusinfluenzainfluenzainfluenzainfluenza andandandand H.H.H.H.

parainfluenzaeparainfluenzaeparainfluenzaeparainfluenzae2222

BlastomycesBlastomycesBlastomycesBlastomycesdermatitidisdermatitidisdermatitidisdermatitidis 3333

NeisseriaNeisseriaNeisseriaNeisseriagonorrhaeaegonorrhaeaegonorrhaeaegonorrhaeae 4444

MycobacteriumMycobacteriumMycobacteriumMycobacterium

tuberculosistuberculosistuberculosistuberculosis5555

UNCOMMONUNCOMMONUNCOMMONUNCOMMON UROPATHOGENSUROPATHOGENSUROPATHOGENSUROPATHOGENSM.TUBERCULOSIS PRODUCES CHRONIC INFECTIONM.TUBERCULOSIS PRODUCES CHRONIC INFECTIONM.TUBERCULOSIS PRODUCES CHRONIC INFECTIONM.TUBERCULOSIS PRODUCES CHRONIC INFECTION

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PATHOPHYSIOLOGY OF URINARY PATHOPHYSIOLOGY OF URINARY PATHOPHYSIOLOGY OF URINARY PATHOPHYSIOLOGY OF URINARY TRACT INFECTIONTRACT INFECTIONTRACT INFECTIONTRACT INFECTION

Ascending route of infection most common

Colonization of urethra and periurethral tissue byuropathogens the initial event in urinary tract infectionUrinary tract infection more common in women than men dueto short female urethra with distention and turbulent flow that

washes urethral organisms into the bladder during micturitionand in close proximity to perianal areasHospital infection associated with lower urinary tractinstrumentation (catheterization, cystoscopy)Once in the bladder uropathogens multiply, then pass up theureters (especially if vesicoureteral reflux present) to therenal pelvis and parenchymaSource of uropathogens: enteric bacteria

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There are several different methods for collectionof a urine sample. The most common is themidstream cleanmidstream cleanmidstream cleanmidstream clean- ---catch techniquecatch techniquecatch techniquecatch technique . Hands shouldbe washed before beginning. For females, the

external genitalia (sex organs) are washed two orthree times with a cleansing agent and rinsed withwater. In males, the external head of the penis is

similarly cleansed and rinsed. The patient is theninstructed to begin to urinate, and the urine iscollected midstream into a sterile container.

COLLECTING THE URINE

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The urine collected in a

wide mouthed containerfrom patientsA mid stream specimen isthe most ideal for

processingFemale patients passesurine with a labiaseparated and mid

stream sample iscollected

SPECIMEN COLLECTION

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10

Female patients

Urine collected with InstructionsUrine collected with InstructionsUrine collected with InstructionsUrine collected with Instructions

Spreadlabia,

using plainsoap or antiseptic

wipe front toback,

dry with

tissues

Retractprepuce,

using plainsoap or antisepticclean glans.Dry withtissues.

Male patientsBegin passing urineStop flow in midstream

Pass several ml into pen container without touching rim

Stop flow before it ends

Recap container

Pass remaining urine into lavatory

Send specimen to laboratoryimmediately

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Urine collected in s terileUrine collected in sterileUrine collected in s terileUrine collected in sterile

specimen container mustspecimen container mustspecimen container mustspecimen container mustbe processedbe processedbe processedbe processed within 2within 2within 2within 2hours, or refrigerated andhours, or refrigerated andhours, or refrigerated andhours, or refrigerated andprocessed within 24 hoursprocessed within 24 hoursprocessed within 24 hoursprocessed within 24 hoursUrine collected in s terileUrine collected in sterileUrine collected in s terileUrine collected in sterilespecimen container withspecimen container withspecimen container withspecimen container withborateborateborateborate preservativepreservativepreservativepreservativeshould be processedshould be processedshould be processedshould be processedwithin 24 hours (nowithin 24 hours (nowithin 24 hours (nowithin 24 hours (no

refrigeration required)refrigeration required)refrigeration required)refrigeration required)

COLLECTIONCOLLECTIONCOLLECTIONCOLLECTION AND PRESERVATION OFAND PRESERVATION OFAND PRESERVATION OFAND PRESERVATION OF URINEURINEURINEURINESPECIMENSSPECIMENSSPECIMENSSPECIMENS

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In infants, a urinarycollection bag (plasticbag with an adhesiveseal on one end) is

attached over thelabia in girls or a boy'spenis to collect the

specimen.

COLLECTING URINE IN INFANTS AND YOUNGCOLLECTING URINE IN INFANTS AND YOUNGCOLLECTING URINE IN INFANTS AND YOUNGCOLLECTING URINE IN INFANTS AND YOUNGCHILDREN DIFFERS FROM ADULTSCHILDREN DIFFERS FROM ADULTSCHILDREN DIFFERS FROM ADULTSCHILDREN DIFFERS FROM ADULTS

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Non invasive methods are safeand idealFollow the Broomhall et almethodBy tapping just above thepubis with two fingers place

on suprapubic region after 1hour of feed, tapping on atthe rate of 1 tap/second for aperiod of 1 minute, if notsuccessful tapping is repeated

once aging.The child spontaneo usly pass the

Urine and to be col lected in aster i le container

SPECIMEN COLLECTION IN INFANTSSPECIMEN COLLECTION IN INFANTSSPECIMEN COLLECTION IN INFANTSSPECIMEN COLLECTION IN INFANTSAND YOUNG CHILDRENAND YOUNG CHILDRENAND YOUNG CHILDRENAND YOUNG CHILDREN

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Another method is the

catheterized urinespecimen in which alubricated catheter (thinrubber tube) is inser tedthrough the urethra (tube-

like structure in whichurine is expelled from thebladder) into the bladder.This avoids contaminationfrom the urethra orexternal genitalia.

CATHETERIZED URINECATHETERIZED URINECATHETERIZED URINECATHETERIZED URINE

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If the patient already has aurinary catheter in place, aurine specimen may becollected by clamping thetubing below the collectionport and using a sterileneedle and syringe toobtain the urine sample;urine cannot be taken fromthe drainage bag, as it isnot fresh and has had anopportunity to grow bacteriaat room temperatur

SPECIMEN CAN BE COLLECTED AFTER CLAMPINGTHE CATHETER

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On rare occasions, the

health care providermay collect a urinesample by inserting aneedle directly into thebladder (suprapubic tap)and draining the urine;this method is used onlywhen a sample isneeded quickly andtechnically competentstaff are available

SUPRAPUBIC PUNCTURE ASUPRAPUBIC PUNCTURE ASUPRAPUBIC PUNCTURE ASUPRAPUBIC PUNCTURE ATECHNICALLY DEMANDING METHODTECHNICALLY DEMANDING METHODTECHNICALLY DEMANDING METHODTECHNICALLY DEMANDING METHOD

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If delivery of the urine

specimen to thelaboratory within onehour of collection is notpossible, it should berefrigerated. The healthcare provider should beinformed of anyantibiotics currently orrecently taken.

SENDING THE SPECIMEN TO LABORATORY SENDING THE SPECIMEN TO LABORATORY SENDING THE SPECIMEN TO LABORATORY SENDING THE SPECIMEN TO LABORATORY

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DIAGNOSIS OF URINARY TRACTDIAGNOSIS OF URINARY TRACTDIAGNOSIS OF URINARY TRACTDIAGNOSIS OF URINARY TRACTINFECTIONINFECTIONINFECTIONINFECTION

Step 1Microscopy of Urine fordetection of Pyuria,

Leucocytes should befound in numbers of atleast as great as 10 4 /ml before the pyuria isestablished

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WET FILM EXAMINATION OF URINEWET FILM EXAMINATION OF URINEWET FILM EXAMINATION OF URINEWET FILM EXAMINATION OF URINEAll wet films to be examinedwith high power ( x 40 )

objective.Prepare the drop of urine aftermixing the urine withoutcentrifugationTransfer 0.05 ml on th e middle

of the microscope slide andcover slip is applied.The prepared specimen show asmall excess of fluid along theedges of the cover slip.A approximate finding of 1leukocyte / 7 high power fieldscorresponds to presence ofpyuria.

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Plate: provide large

surface for isolation andobservation of coloniesUsing a sterile loop or asterile swab streak your

sample on the petri plateImpor tant let yoursterilized loop cool beforeyou pick up your sample

INOCULATING THE CULTURE PLATE WITHURINE


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SELECTION OF LOOP FORSELECTION OF LOOP FORSELECTION OF LOOP FORSELECTION OF LOOP FORSEMIQUANTITATIVE METHODSEMIQUANTITATIVE METHODSEMIQUANTITATIVE METHODSEMIQUANTITATIVE METHOD

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SPECIMEN INOCULATIONSAll cultures processed bySemiquantitative method a loop

of standard dimension ofapproximately known volume isinoculated into selected cultureplateIn gene ral a loop of SWG 28with a diameter of 3.26 mminternal diameter which canhold a drop of water or urine0.004 ml.After inoculation the cultureplates are incubated at 37 0 cextending to > 18 hours arereadThe colony counts are made, aseach colony corropsdes tonumber of viable bacteria per

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CULTURING OF URINE FOR ISOLATIONBACTERIAL PATHOGENS

SemiquantitativeSemiquantitativeSemiquantitativeSemiquantitative

culturecultureculturecultureSelect the MediaFor common isolatesMac Conkeys agar

helps in differentiationof Lactose fermentingorganisms from non

lactose fermentingpathogens

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MacConkey agar

utilized as selectivedifferential agar for gram-negativebacteria,Colistin nalidixicacid agar asselective agar for gram-positivebacteria, and

chocolate agar for fastidious gram-negative bacteria(Haemophilus )

CHOOSING MEDIA TO SUITCHOOSING MEDIA TO SUITCHOOSING MEDIA TO SUITCHOOSING MEDIA TO SUITMICROORGANISMS IS IMPORTANTMICROORGANISMS IS IMPORTANTMICROORGANISMS IS IMPORTANTMICROORGANISMS IS IMPORTANT

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Inoculation of urine for

quantitative culture(colony formingunitscfus) performedwith a calibrated 0.001

mL and 0.01 mL plasticor wire loopSheep blood agar (SBA)utilized for quantitativeurine culture

INOCULATION OF URINEINOCULATION OF URINEINOCULATION OF URINEINOCULATION OF URINE

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CULTURE MEDIA FOR ISOLATIONSCULTURE MEDIA FOR ISOLATIONSCULTURE MEDIA FOR ISOLATIONSCULTURE MEDIA FOR ISOLATIONS

CLEDCLEDCLEDCLED MEDIUMMEDIUMMEDIUMMEDIUMI t is also an excellent universalI t is also an excellent universalI t is also an excellent universalI t is also an excellent universal

culture medium owing to itsculture medium owing to itsculture medium owing to itsculture medium owing to itswide spectrum of nutrients,wide spectrum of nutrients,wide spectrum of nutrients,wide spectrum of nutrients,lack of i nhibitors and the factlack of i nhibitors and the factlack of i nhibitors and the factlack of i nhibitors and the factthat i t al lows a cer tain degreethat i t al lows a cer tain degreethat i t al lows a cer tain degreethat i t al lows a cer tain degreeor differentiat ion between theor differentiat ion between theor differentiat ion between theor differentiat ion between thecolonies. I t contains lactose ascolonies. I t contains lactose ascolonies. I t contains lactose ascolonies. I t contains lactose asa reactive compound which,a reactive compound which,a reactive compound which,a reactive compound which,when degraded to acid, causeswhen degraded to acid, causeswhen degraded to acid, causeswhen degraded to acid, causes

bromothymol blue to change itsbromothymol blue to change itsbromothymol blue to change itsbromothymol blue to change itscolour to yellow. Alkalinizationcolour to yellow. Alkalinizationcolour to yellow. Alkalinizationcolour to yellow. Alkalinizationproduces a deep blueproduces a deep blueproduces a deep blueproduces a deep bluecolourat ion. The lac k ofcolourat ion. The l ack ofcolourat ion. The lac k ofcolourat ion. The l ack ofelectrolytes suppresses theelectrolytes suppresses theelectrolytes suppresses theelectrolytes suppresses theswarming of Proteusswarming of Proteusswarming of Proteusswarming of ProteusLaboratories which haveLaboratories which haveLaboratories which haveLaboratories which havedifficu lty in availability of diff iculty in availability of difficu lty in availability of diff iculty in availability of blood agar, CLED medium isblood agar, CLED medium isblood agar, CLED medium isblood agar, CLED medium isoptedoptedoptedopted

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ENRICHED CULTURE MEDIA FORENRICHED CULTURE MEDIA FORENRICHED CULTURE MEDIA FORENRICHED CULTURE MEDIA FORISOLATIONISOLATIONISOLATIONISOLATION

Blood agarBlood agarBlood agarBlood agarhelps in isolation offastidious, extractingstrains May extendedincubation for isolationof pathogens for morethan 48 hours with

added atmosphere of 5 10 % co 2

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READING THE CULTURE PLATESREADING THE CULTURE PLATESREADING THE CULTURE PLATESREADING THE CULTURE PLATES

A true infection in the absence of prior antibiotic therapy the

number of bacteria is likely to be at least 101010105555

or more.Contaminated specimens present with colony counts 10 4 to be considered aspresence of Urinary tract infection with the supporting clinicalhistoryOn some occasions more than one pathogen is isolated butshould be processed for all practical purposes

eg E.coli along with Streptococcus fecalisOn few occasions even counts 10 3 are proved

significant

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IDENTIFICATION OF GRAM +IDENTIFICATION OF GRAM +IDENTIFICATION OF GRAM +IDENTIFICATION OF GRAM +ORGANISMSORGANISMSORGANISMSORGANISMS

All colonies identifiedmorphologically as

Staphylococcus to becharacterized as

Staphylococcus aureusStaphylococcussaprophyticus

Staphylococcusepidermidis

Enterococci - fecal group of organisms

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With 0.001 ml loop,1 colony on SBAequivalent to 1,000cfus per mL of urineWith 0.01 ml loop,1 colony on SBAequivalent to 100cfus per mL of urine

COUNTING THE COLONIES

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A single species of Enterobacter iaceae

recovered at >10 5 cfus/mLurine: with patientssymptomatic for urinary tractinfection, 95% probability of true bacteriuria

A single species of Enterobacter iaceaerecovered at 10 4 -10 5 cfus/mLurine: with patientssymptomatic for urinary tractinfection, 33% probability of true bacteriuria

INTERPRETATION OFINTERPRETATION OFINTERPRETATION OFINTERPRETATION OFENTEROBACTERIACEAEENTEROBACTERIACEAEENTEROBACTERIACEAEENTEROBACTERIACEAE

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A single species of Enterobacteriaceae

recovered at >105

cfus/mL urine: withpatients symptomatic for urinary tract infection,95% probability of truebacteriuria

A single species of Enterobacteriaceaerecovered at 10 4-10 5cfus/mL urine: withpatients symptomatic for urinary tract infection,

33% probability of truebacteriuria

INTERPRETATION OF ENTEROBACTERIACEAEINTERPRETATION OF ENTEROBACTERIACEAEINTERPRETATION OF ENTEROBACTERIACEAEINTERPRETATION OF ENTEROBACTERIACEAE

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Significant bacteriuria in anasymptomatic patient is100,000 or more colonies permilli l i ter of urine from amidstream, clean-catchspecimen; yet, a colony count

of 200 Escherichia coli per mlmay be significant in amidstream male void orcatheterized female. About95% of all positive UTIcultures will produceessentially pure cultures ifurine is colle cted carefully andthe media inoculatedpromptly.

WHAT ISWHAT ISWHAT ISWHAT IS SIGNIFICANT BACTERIURIASIGNIFICANT BACTERIURIASIGNIFICANT BACTERIURIASIGNIFICANT BACTERIURIA

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CONCEPT OF SIGNIFICANT BACTERIURIACONCEPT OF SIGNIFICANT BACTERIURIACONCEPT OF SIGNIFICANT BACTERIURIACONCEPT OF SIGNIFICANT BACTERIURIA

Up to 10 4 /ml considered normal

i .e . Insignif icant

10 5 /ml and above considere d to

be Signif icant

Concept val id only for voided

specimen of urine

Exceptions - s low growing

organisms, pat ient on ant ibiot ictherapy, diuret ic therapy

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A single species of

Enterobacteriaceaerecovered at 10 4 -10 5

cfus/mL urine: with

patients symptomaticfor urinary tractinfection, 33%

probability of truebacteriuria

WHAT CAN BE A SIGNIFICANT COUNTWHAT CAN BE A SIGNIFICANT COUNTWHAT CAN BE A SIGNIFICANT COUNTWHAT CAN BE A SIGNIFICANT COUNT

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Gram-positive, fungal,

and fastidiousuropathogens oftenpresent in lowernumbers (10 4 -10 5

cfus/mL urine)Urethral commensalsrecovered at


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IDENTIFICATION OF ISOLATESIDENTIFICATION OF ISOLATESIDENTIFICATION OF ISOLATESIDENTIFICATION OF ISOLATESGRAM + ISOLATESGRAM + ISOLATESGRAM + ISOLATESGRAM + ISOLATES

The minimal tests todifferentiate Gram +cocci include

1 Catalase1 Catalase1 Catalase1 Catalase2 Coagulase test3 Bile esculin testing4 Bacitracin in

Streptococcus isolates

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BIOCHEMICAL TESTS IN GRAM - VE BACILLI

Catalase testOxidase testNitrite reduction testIndole test

Methyl red testV P testCitrate test

Decarboxylation testsLysine, ornithine, Arginine

tests

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39Dr.T.V.Rao MD

PEER REVIEWED URINECULTURE

INTERPRETATIONS


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Routine : uncomplicated urinary tract infection inambulatory outpatients (0.001 mL loop, SBA,MAC; 24 hr. incubation)Surveillance : neurogenic bladder, indwellingcatheter, geriatric patents (0.001 mL loop, SBA,MAC, CNA; 24 hr. incubation)Special : suprapubic aspirates or straight catheterspecimens where previous cultures negative,unresponsive to therapy, or possibili ty of unusualurinary tract pathogen (0.001 and 0.01 mL loop,

BA, MAC, CHOC; minimum 48 hr. incubation1111 Clarridge, Johnson, Pezzlo, an d Weissfeld, ASM Cumitech 2B,Clarridge, Johnson, Pe zzlo, and Weissfeld, ASM Cumitech 2B,Clarridge, Johnson, Pezzlo, an d Weissfeld, ASM Cumitech 2B,Clarridge, Johnson, Pe zzlo, and Weissfeld, ASM Cumitech 2B,November 1998.November 1998.November 1998.November 1998.

CUMITECH GUIDELINESCUMITECH GUIDELINESCUMITECH GUIDELINESCUMITECH GUIDELINES FOR INOCULATION OFFOR INOCULATION OFFOR INOCULATION OFFOR INOCULATION OFURINE CULTURESURINE CULTURESURINE CULTURESURINE CULTURES 1111

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One isolate at >10 4:

Full ID andSusceptibilityOne or two gram-negative isolates at >10 5and other isolates atleast 10X less : Full ID

and Susceptibility of gram-negative isolates

CUMITECH GUIDELINESCUMITECH GUIDELINESCUMITECH GUIDELINESCUMITECH GUIDELINES FOR INTERPRETATION OFFOR INTERPRETATION OFFOR INTERPRETATION OFFOR INTERPRETATION OFROUTINE URINE CULTURESROUTINE URINE CULTURESROUTINE URINE CULTURESROUTINE URINE CULTURES 1111

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Other patterns of

isolates at >104

:Presumptive IDonly

Ignore mixedurethral flora at


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One or twoisolates at >10 2to 10 5 : Full ID andSusceptibility is

essential beforeconfirmed asuncommon

isolates

CUMITECH GUIDELINESCUMITECH GUIDELINESCUMITECH GUIDELINESCUMITECH GUIDELINES FOR INTERPRETATION OFFOR INTERPRETATION OFFOR INTERPRETATION OFFOR INTERPRETATION OF

SPECIAL OR UNCOMMONSPECIAL OR UNCOMMONSPECIAL OR UNCOMMONSPECIAL OR UNCOMMON URINE CULTURESURINE CULTURESURINE CULTURESURINE CULTURES 1111

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Midstream, female with cystitis, >10 2 with positive urineleukocyte esteraseMidstream, female with pyelonephritis, >10 5 with positiveurine leukocyte esteraseMidstream, asymptomatic, >10 5 with negative urineleukocyte esterase (usually)Midstream, male with UTI: >10 3 with leukocyte with urineleukocyte esterase positiveStraight catheter: >10 2 with urine leukocyte esterasepositiveIndwelling catheter: >10 3 with urine leukocyte esterase

positive or negative1111 Manual of Clinical Microbiology, 8Manual of Clinical Microbiology, 8Manual of Clinical Microbiology, 8Manual of Clinical Microbiology, 8 thththth Edition, ASM, 2003Edition, ASM, 2003Edition, ASM, 2003Edition, ASM, 2003

ASM MANUAL GUIDELINES FOR URINE CULTUREASM MANUAL GUIDELINES FOR URINE CULTUREASM MANUAL GUIDELINES FOR URINE CULTUREASM MANUAL GUIDELINES FOR URINE CULTURE

RESULTS LIKELY TO BE SIGNIFICANTRESULTS LIKELY TO BE SIGNIFICANTRESULTS LIKELY TO BE SIGNIFICANTRESULTS LIKELY TO BE SIGNIFICANT 1111

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Urine leukocyte esterase positiveUrine leukocyte esterase positiveUrine leukocyte esterase positiveUrine leukocyte esterase positive

OneOneOneOne or two organisms at >10or two organisms at >10or two organisms at >10or two organisms at >10 3333 : Full: Full: Full: Full IDIDIDID andandandandSusceptibilitySusceptibilitySusceptibilitySusceptibilityOneOneOneOne organism at >10organism at >10organism at >10organism at >10 4444 with others (2 orwith others (2 orwith others (2 orwith others (2 or

moremoremoremore) at least 10X less: Full ID and) at least 10X less: Full ID and) at least 10X less: Full ID and) at least 10X less: Full ID andsusceptibilitysusceptibilitysusceptibilitysusceptibility of predominantof predominantof predominantof predominant organismorganismorganismorganismReportReportReportReport all group Ball group Ball group Ball group B ----hemolytichemolytichemolytichemolytic streptococcistreptococcistreptococcistreptococci

for women < 50 yearsfor women < 50 yearsfor women < 50 yearsfor women < 50 years1111 Modified from ASM Cumitech, ASM Manual, and CDCModified from ASM Cumitech, ASM Manual, and CDCModified from ASM Cumitech, ASM Manual, and CDCModified from ASM Cumitech, ASM Manual, and CDCMMWR 2002;51 (RRMMWR 2002;51 (RRMMWR 2002;51 (RRMMWR 2002;51 (RR- ---11):111):111):111):1- ---22222222

NMH GUIDELINES FOR INTERPRETATION OFNMH GUIDELINES FOR INTERPRETATION OFNMH GUIDELINES FOR INTERPRETATION OFNMH GUIDELINES FOR INTERPRETATION OFURINE CULTURESURINE CULTURESURINE CULTURESURINE CULTURES 1111

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NMH GUIDELINESNMH GUIDELINESNMH GUIDELINESNMH GUIDELINES FOR INTERPRETATION OFFOR INTERPRETATION OFFOR INTERPRETATION OFFOR INTERPRETATION OFURINE CULTURESURINE CULTURESURINE CULTURESURINE CULTURES 1111

Urine leukocyte esterase negativeOne or two organisms at >105: Full andSusceptibilityOne gram-negative organism (pure culture) at>104: Full ID and SusceptibilityYeast in pure culture: ID asCandida albicans or notC. albicansReport all group B -hemolytic streptococci for

women


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ANTIBIOTIC SENSITIVITY TESTING

All the isolatedbacteria identifiedas pathogenic to betested for AntibioticSensitivity/Resistance

pattern by diskdiffusion methods

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ANTIBIOTIC SENSITIVITY ANTIBIOTIC SENSITIVITY ANTIBIOTIC SENSITIVITY ANTIBIOTIC SENSITIVITY Always use a pure

growth of the isolate.Perform testing forantibiotic sensitivitywith inoculum whichis comparable todefined McFarlandMcFarlandMcFarlandMcFarlandstandards

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High potency disks

usedAntibiotics normallyactive in urineselectedAntibiotics found inhigher concentrationin urine preferredPrimary sensitivity testfrequently used

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ANTIBIOTIC SENSITIVITY TESTINGANTIBIOTIC SENSITIVITY TESTINGANTIBIOTIC SENSITIVITY TESTINGANTIBIOTIC SENSITIVITY TESTING

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Microscopy helps todetect pus cells andepithelial cells, aninsignificant growth withfew pus cells can ignoredas contaminants

In females even thepresence of leucocyteswith Squamous epithelialcells without a definedsignificant growth shouldbe ignored.

REPORTING OF CONTAMINATEDREPORTING OF CONTAMINATEDREPORTING OF CONTAMINATEDREPORTING OF CONTAMINATEDSPECIMENSSPECIMENSSPECIMENSSPECIMENS

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Negative culture

results showing nobacterial growth areavailable after 24hours. Positive resultsrequire 24-72 hours tocompleteidentification of the

number and type ofbacteria found

REPORTING OF NEGATIVE URINEREPORTING OF NEGATIVE URINEREPORTING OF NEGATIVE URINEREPORTING OF NEGATIVE URINECULTURESCULTURESCULTURESCULTURES

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CLINICAL PROBLEMS MANIFESTING AS


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CLINICAL PROBLEMS MANIFESTING AS

URINARY TRACT INFECTIONS

An infection of theGenitourinary tractwhere the MicrobesDonot grow on selectedmedia as inMycobacteriumtuberculosis

Gonococcus

Nutritionally exactingor anaerobic bacteriashould be considered.

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Most over diagnosedcondition, both by theclinicians andmicrobiologists.Makes the physicians tomiss the existingcondition.Unnecessary antibioticprescription lead todrug resistance, loss ofresources

URINARY TRACT INFECTION ARE OVERURINARY TRACT INFECTION ARE OVERURINARY TRACT INFECTION ARE OVERURINARY TRACT INFECTION ARE OVER

DIAGNOSEDDIAGNOSEDDIAGNOSEDDIAGNOSED

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Dr.T.V.Rao MD 54

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Culturing of Urine Skills in Microbiology

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