CULTURING OF URINE SKILL BASED LEARNING         Dr.T.V.Rao.MD              Dr.T.V.Rao MD              1
OVER VIEW OF URINARY TRACT           INFECTIONSMost common infectious diseaseMost Numerous specimens are received in the L...
WHAT IS URINARY TRACT INFECTIONan infection of one ormore structures in theurinary system. Most UTIsare caused by gram-neg...
URINARY TRACT INFECTIONS ARE MORE        COMMON IN WOMEN                         The condition is more                    ...
COMMON UROPATHOGENS Escherichia coli                            Klebsiella, Enterobacter, Other Enterobacteriaceae ( Klebs...
UNCOMMON UROPATHOGENS M.TUBERCULOSIS PRODUCES CHRONIC INFECTIONCorynebacteriumurealyticum 1Haemophilusinfluenza and H.para...
PATHOPHYSIOLOGY OF URINARY        TRACT INFECTIONAscending route of infection most commonColonization of urethra and periu...
COLLECTING THE URINEThere are several different methods for collectionof a urine sample. The most common is themidstream c...
SPECIMEN COLLECTIONThe urine collected in awide mouthed containerfrom patientsA mid stream specimen isthe most ideal forpr...
Urine collected with InstructionsFemale patients                                                Male patients             ...
COLLECTION AND PRESERVATION OF URINE             SPECIMENSUrine collected in sterilespecimen container mustbe processed wi...
COLLECTING URINE IN INFANTS AND YOUNG    CHILDREN DIFFERS FROM ADULTSIn infants, a urinarycollection bag (plasticbag with ...
SPECIMEN COLLECTION IN INFANTS     AND YOUNG CHILDREN Non invasive methods are safe and ideal Follow the Broomhall et al m...
CATHETERIZED URINE                  Another method is the                  catheterized urine                  specimen in...
SPECIMEN CAN BE COLLECTED AFTER CLAMPING              THE CATHETERIf the patient already has aurinary catheter in place, a...
SUPRAPUBIC PUNCTURE ATECHNICALLY DEMANDING METHOD                       On rare occasions, the                       healt...
SENDING THE SPECIMEN TO LABORATORYIf delivery of the urinespecimen to thelaboratory within onehour of collection is notpos...
DIAGNOSIS OF URINARY TRACT           INFECTIONStep 1Microscopy of Urine fordetection of Pyuria,Leucocytes should befound i...
WET FILM EXAMINATION OF URINEAll wet films to be examinedwith high power ( x 40 )objective.Prepare the drop of urine after...
INOCULATING THE CULTURE PLATE WITH               URINEPlate: provide largesurface for isolation andobservation of colonies...
SELECTION OF LOOP FORSEMIQUANTITATIVE METHOD          Dr.T.V.Rao MD   21
SPECIMEN INOCULATIONS               All cultures processed by                     Semiquantitative method a loop          ...
CULTURING OF URINE FOR ISOLATION OF       BACTERIAL PATHOGENSSemiquantitativecultureSelect the MediaFor common isolates Ma...
CHOOSING MEDIA TO SUIT  MICROORGANISMS IS IMPORTANTMacConkey agarutilized as selectivedifferential agar forgram-negativeba...
INOCULATION OF URINEInoculation of urine forquantitative culture(colony formingunits→cfu’s) performedwith a calibrated 0.0...
CULTURE MEDIA FOR ISOLATIONS       CLED MEDIUM                        It is also an excellent universal                   ...
ENRICHED CULTURE MEDIA FOR            ISOLATIONBlood agar helps in isolation offastidious, extractingstrains May extendedi...
READING THE CULTURE PLATES A true infection in the absence of prior antibiotic therapy the number of bacteria is likely to...
IDENTIFICATION OF GRAM +             ORGANISMS  All colonies identified   morphologically as  Staphylococcus to be     cha...
COUNTING THE COLONIES                   With 0.001 ml loop,                   1 colony on SBA                   equivalent...
INTERPRETATION OF             ENTEROBACTERIACEAEA single species ofEnterobacteriaceaerecovered at >10 5 cfu’s/mLurine: wit...
INTERPRETATION OF ENTEROBACTERIACEAE A single species of Enterobacteriaceae recovered at >10 5 cfu’s/mL urine: with patien...
WHAT IS SIGNIFICANT BACTERIURIA Significant bacteriuria in anasymptomatic patient is100,000 or more colonies permilliliter...
CONCEPT OF SIGNIFICANT BACTERIURIAUp to 10 4 /ml considered normali.e. Insignificant10 5 /ml and above considered tobe Sig...
WHAT CAN BE A SIGNIFICANT COUNT                        A single species of                        Enterobacteriaceae      ...
GRAM POSITIVES AND FUNGI THE COUNTS            MAY BE <10 5Gram-positive, fungal,and fastidiousuropathogens oftenpresent i...
IDENTIFICATION OF ISOLATES          GRAM + ISOLATESThe minimal tests todifferentiate Gram +cocci include 1 Catalase 2 Coag...
BIOCHEMICAL TESTS IN  GRAM - VE BACILLI                   Catalase test                   Oxidase test                   N...
PEER REVIEWED URINE           CULTURE    INTERPRETATIONS          Dr.T.V.Rao MD   39
CUMITECH GUIDELINES FOR INOCULATION OF           URINE CULTURES 1 Routine: uncomplicated urinary tract infection in ambula...
CUMITECH GUIDELINES FOR INTERPRETATION OF         ROUTINE URINE CULTURES 1One isolate at >10 4:Full ID andSusceptibilityOn...
CUMITECH GUIDELINES FOR INTERPRETATION OF         ROUTINE URINE CULTURES 1                             Other patterns of  ...
CUMITECH GUIDELINES FOR INTERPRETATION OF  SPECIAL OR UNCOMMON URINE CULTURES 1 One or two isolates at >10 2 to 10 5: Full...
ASM MANUAL GUIDELINES FOR URINE CULTURE     RESULTS LIKELY TO BE SIGNIFICANT 1  Midstream, female with cystitis, >10 2 wit...
NMH GUIDELINES FOR INTERPRETATION OF           URINE CULTURES 1 Urine leukocyte esterase positive One or two organisms at ...
NMH GUIDELINES FOR INTERPRETATION OF          URINE CULTURES 1Urine leukocyte esterase negativeOne or two organisms at >10...
ANTIBIOTIC SENSITIVIT Y TESTINGAll the isolatedbacteria identifiedas pathogenic to betested for AntibioticSensitivity/Resi...
ANTIBIOTIC SENSITIVITYAlways use a puregrowth of the isolate.Perform testing forantibiotic sensitivitywith inoculum whichi...
ANTIBIOTIC SENSITIVITY TESTINGHigh potency disksusedAntibiotics normallyactive in urineselectedAntibiotics found inhigher ...
REPORTING OF CONTAMINATED          SPECIMENSMicroscopy helps todetect pus cells andepithelial cells, aninsignificant growt...
REPORTING OF NEGATIVE URINE            CULTURESNegative cultureresults showing nobacterial growth areavailable after 24hou...
CLINICAL PROBLEMS MANIFESTING AS    URINARY TRACT INFECTIONSAn infection of theGenitourinary tractwhere the MicrobesDonot ...
URINARY TRACT INFECTION ARE OVER           DIAGNOSED                        Most over diagnosed                        con...
Created by Dr.T.V.Rao MD for       ‘eLearning” resources forMicrobiologists in the Developing World                  Email...
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Culturing of urine, Skill based learning

  1. 1. CULTURING OF URINE SKILL BASED LEARNING Dr.T.V.Rao.MD Dr.T.V.Rao MD 1
  2. 2. OVER VIEW OF URINARY TRACT INFECTIONSMost 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 infor mation gives manyclues for better diagnostic evaluations.Specimen collection is the primary objective in getting anideal sample. Dr.T.V.Rao MD 2
  3. 3. WHAT IS URINARY TRACT INFECTIONan infection of one ormore 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 Dr.T.V.Rao MD 3
  4. 4. URINARY TRACT INFECTIONS ARE MORE COMMON IN WOMEN The condition is more common in women than in men. UTI may be asymptomatic but is usually characterized by urinary frequency, burning pain with voiding, and, if the infection is severe, visible blood and pus in the urine. Fever and back pain often accompany kidney infections Dr.T.V.Rao MD 4
  5. 5. COMMON UROPATHOGENS Escherichia coli Klebsiella, Enterobacter, Other Enterobacteriaceae ( Klebsiella , Enterobacter , Proteus, Citrobacter) Proteus , Citrobacter ) Pseudomonas aeruginosa Enterococcus Staphylococcus saprophyticus Staphylococcus aureus Streptococcus agalactiae (group B) CandidaAssociated with staphylococcemiaDenotes vaginal colonization in pregnant women Dr.T.V.Rao MD 5
  6. 6. UNCOMMON UROPATHOGENS M.TUBERCULOSIS PRODUCES CHRONIC INFECTIONCorynebacteriumurealyticum 1Haemophilusinfluenza and H.parainfluenzae 2Blastomycesdermatitidis 3Neisseriagonorrhaeae 4Mycobacteriumtuberculosis 5 Dr.T.V.Rao MD 6
  7. 7. PATHOPHYSIOLOGY OF URINARY TRACT INFECTIONAscending route of infection most commonColonization 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 thatwashes 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 Dr.T.V.Rao MD 7
  8. 8. COLLECTING THE URINEThere are several different methods for collectionof a urine sample. The most common is themidstream clean - catch technique Hands should clean- technique.be washed before beginning. For females, theexternal genitalia (sex organs) are washed two orthree times with a cleansing agent and rinsed withwater. In males, the external head of the penis issimilarly cleansed and rinsed. The patient is theninstructed to begin to urinate, and the urine iscollected midstream into a sterile container. Dr.T.V.Rao MD 8
  9. 9. SPECIMEN COLLECTIONThe urine collected in awide mouthed containerfrom patientsA mid stream specimen isthe most ideal forprocessingFemale patients passesurine with a labiaseparated and midstream sample iscollected Dr.T.V.Rao MD 9
  10. 10. Urine collected with InstructionsFemale patients Male patients Begin passing urineSpread Stop flow in midstream Retractlabia, prepuce, Pass several ml into pen container without touching rim using plainusing plain soap orsoap or Stop flow before it ends antisepticantiseptic Recap container clean glans.wipe front to Pass remaining urine into lavatory Dry withback, tissues. Send specimen to laboratorydry with immediatelytissues (refrigerate if prolonged transport time) Dr.T.V.Rao MD 10
  11. 11. COLLECTION AND PRESERVATION OF URINE SPECIMENSUrine collected in sterilespecimen container mustbe processed within 2hours, or refrigerated andprocessed within 24 hoursUrine collected in sterilespecimen container withborate preservativeshould be processedwithin 24 hours (norefrigeration required) Dr.T.V.Rao MD 11
  12. 12. COLLECTING URINE IN INFANTS AND YOUNG CHILDREN DIFFERS FROM ADULTSIn infants, a urinarycollection bag (plasticbag with an adhesiveseal on one end) isattached over thelabia in girls or a boyspenis to collect thespecimen. Dr.T.V.Rao MD 12
  13. 13. SPECIMEN COLLECTION IN INFANTS AND YOUNG CHILDREN Non invasive methods are safe and ideal Follow the Broomhall et al method By tapping just above the pubis with two fingers place on suprapubic region after 1 hour of feed, tapping on at the rate of 1 tap/second for a period of 1 minute, if not successful tapping is repeated once aging.The child spontaneously pass the Urine and to be collected in a sterile container Dr.T.V.Rao MD 13
  14. 14. CATHETERIZED URINE Another method is the catheterized urine specimen in which a lubricated catheter (thin rubber tube) is inserted through the urethra (tube- like structure in which urine is expelled from the bladder) into the bladder. This avoids contamination from the urethra or external genitalia. Dr.T.V.Rao MD 14
  15. 15. SPECIMEN CAN BE COLLECTED AFTER CLAMPING THE CATHETERIf 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 Dr.T.V.Rao MD 15
  16. 16. SUPRAPUBIC PUNCTURE ATECHNICALLY DEMANDING METHOD On rare occasions, the health care provider may collect a urine sample by inserting a needle directly into the bladder (suprapubic tap) and draining the urine; this method is used only when a sample is needed quickly and technically competent staff are available Dr.T.V.Rao MD 16
  17. 17. SENDING THE SPECIMEN TO LABORATORYIf delivery of the urinespecimen to thelaboratory within onehour of collection is notpossible, it should berefrigerated. The healthcare provider should beinformed of anyantibiotics currently orrecently taken. Dr.T.V.Rao MD 17
  18. 18. DIAGNOSIS OF URINARY TRACT INFECTIONStep 1Microscopy of Urine fordetection of Pyuria,Leucocytes should befound in numbers of atleast as great as 10 4 /ml before the pyuria isestablished Dr.T.V.Rao MD 18
  19. 19. WET 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 the middleof 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. Dr.T.V.Rao MD 19
  20. 20. INOCULATING THE CULTURE PLATE WITH URINEPlate: provide largesurface for isolation andobservation of coloniesUsing a sterile loop or asterile swab streak yoursample on the petri plateImportant let yoursterilized loop cool beforeyou pick up your sample
  21. 21. SELECTION OF LOOP FORSEMIQUANTITATIVE METHOD Dr.T.V.Rao MD 21
  22. 22. SPECIMEN INOCULATIONS All cultures processed by Semiquantitative method a loop of standard dimension of approximately known volume is inoculated into selected culture plate In general a loop of SWG – 28 with a diameter of 3.26 mm internal diameter which can hold a drop of water or urine 0.004 ml. After inoculation the culture plates are incubated at 37 0 c extending to > 18 hours are read The colony counts are made, as each colony corropsdes to number of viable bacteria per ml of urine Dr.T.V.Rao MD 22
  23. 23. CULTURING OF URINE FOR ISOLATION OF BACTERIAL PATHOGENSSemiquantitativecultureSelect the MediaFor common isolates Mac Conkey’s agarhelps in differentiationof Lactose fermentingorganisms from nonlactose fermentingpathogens Dr.T.V.Rao MD 23
  24. 24. CHOOSING MEDIA TO SUIT MICROORGANISMS IS IMPORTANTMacConkey agarutilized as selectivedifferential agar forgram-negativebacteria,Colistin nalidixicacid agar asselective agar forgram-positivebacteria, andchocolate agar forfastidious gram-negative bacteria(Haemophilus) Dr.T.V.Rao MD 24
  25. 25. INOCULATION OF URINEInoculation of urine forquantitative culture(colony formingunits→cfu’s) performedwith a calibrated 0.001mL and 0.01 mL plasticor wire loopSheep blood agar (SBA)utilized for quantitativeurine culture Dr.T.V.Rao MD 25
  26. 26. CULTURE MEDIA FOR ISOLATIONS CLED MEDIUM It is also an excellent universal culture medium owing to its wide spectrum of nutrients, lack of inhibitors and the fact that it allows a certain degree or differentiation between the colonies. It contains lactose as a reactive compound which, when degraded to acid, causes bromothymol blue to change its colour to yellow. Alkalinization produces a deep blue colouration. The lack of electrolytes suppresses the swarming of Proteus Laboratories which have difficulty in availability of blood agar, CLED medium is opted Dr.T.V.Rao MD 26
  27. 27. ENRICHED CULTURE MEDIA FOR ISOLATIONBlood agar helps in isolation offastidious, extractingstrains May extendedincubation for isolationof pathogens for morethan 48 hours withadded atmosphere of 5– 10 % co 2 Dr.T.V.Rao MD 27
  28. 28. READING THE CULTURE PLATES A true infection in the absence of prior antibiotic therapy the number of bacteria is likely to be at least 10 5 or more. Contaminated specimens present with colony counts <10 4, however even less than 10 3 On several occasions the colonies are diverse species Several studies prove counts >10 4 to be considered as presence of Urinary tract infection with the supporting clinical history On some occasions more than one pathogen is isolated but should be processed for all practical purposes eg E.coli along with Streptococcus fecalisOn few occasions even counts 10 3 are proved significant Dr.T.V.Rao MD 28
  29. 29. IDENTIFICATION OF GRAM + ORGANISMS All colonies identified morphologically as Staphylococcus to be characterized as Staphylococcus aureus Staphylococcus saprophyticus Staphylococcus epidermidisEnterococci - fecal group of organisms Dr.T.V.Rao MD 29
  30. 30. COUNTING THE COLONIES With 0.001 ml loop, 1 colony on SBA equivalent to 1,000 cfu’s per mL of urine With 0.01 ml loop, 1 colony on SBA equivalent to 100 cfu’s per mL of urine Dr.T.V.Rao MD 30
  31. 31. INTERPRETATION OF ENTEROBACTERIACEAEA single species ofEnterobacteriaceaerecovered at >10 5 cfu’s/mLurine: with patientssymptomatic for urinary tractinfection, 95% probability oftrue bacteriuriaA single species ofEnterobacteriaceaerecovered at 10 4 -10 5 cfu’s/mLurine: with patientssymptomatic for urinary tractinfection, 33% probability oftrue bacteriuria Dr.T.V.Rao MD 31
  32. 32. INTERPRETATION OF ENTEROBACTERIACEAE A single species of Enterobacteriaceae recovered at >10 5 cfu’s/mL urine: with patients symptomatic for urinary tract infection, 95% probability of true bacteriuria A single species of Enterobacteriaceae recovered at 10 4 -10 5 cfu’s/mL urine: with patients symptomatic for urinary tract infection, 33% probability of true bacteriuria Dr.T.V.Rao MD 32
  33. 33. WHAT IS SIGNIFICANT BACTERIURIA Significant bacteriuria in anasymptomatic patient is100,000 or more colonies permilliliter of urine from amidstream, clean-catchspecimen; yet, a colony countof 200 Escherichia coli per mlmay be significant in amidstream male void orcatheterized female. About95% of all positive UTIcultures will produceessentially pure cultures ifurine is collected carefully andthe media inoculatedpromptly. Dr.T.V.Rao MD 33
  34. 34. CONCEPT OF SIGNIFICANT BACTERIURIAUp to 10 4 /ml considered normali.e. Insignificant10 5 /ml and above considered tobe SignificantConcept valid only for voidedspecimen of urineExceptions - slow growingorganisms, patient on antibiotictherapy, diuretic therapy Dr.T.V.Rao MD 34
  35. 35. WHAT CAN BE A SIGNIFICANT COUNT A single species of Enterobacteriaceae recovered at 10 4 -10 5 cfu’s/mL urine: with patients symptomatic for urinary tract infection, 33% probability of true bacteriuria Dr.T.V.Rao MD 35
  36. 36. GRAM POSITIVES AND FUNGI THE COUNTS MAY BE <10 5Gram-positive, fungal,and fastidiousuropathogens oftenpresent in lowernumbers (10 4 -10 5cfu’s/mL urine)Urethral commensalsrecovered at <10 4cfu’s/mL urine Dr.T.V.Rao MD 36
  37. 37. IDENTIFICATION OF ISOLATES GRAM + ISOLATESThe minimal tests todifferentiate Gram +cocci include 1 Catalase 2 Coagulase test 3 Bile esculin testing 4 Bacitracin inStreptococcus isolates Dr.T.V.Rao MD 37
  38. 38. BIOCHEMICAL TESTS IN GRAM - VE BACILLI Catalase test Oxidase test Nitrite reduction test Indole test Methyl red test V P test Citrate test Decarboxylation tests Lysine, ornithine, Arginine tests Dr.T.V.Rao MD 38
  39. 39. PEER REVIEWED URINE CULTURE INTERPRETATIONS Dr.T.V.Rao MD 39
  40. 40. CUMITECH GUIDELINES FOR INOCULATION OF URINE CULTURES 1 Routine: uncomplicated urinary tract infection in ambulatory outpatients (0.001 mL loop, SBA, MAC; 24 hr. incubation) Surveillance: neurogenic bladder, indwelling catheter, geriatric patents (0.001 mL loop, SBA, MAC, CNA; 24 hr. incubation) Special: suprapubic aspirates or straight catheter specimens where previous cultures negative, unresponsive to therapy, or possibility of unusual urinary tract pathogen (0.001 and 0.01 mL loop, BA, MAC, CHOC; minimum 48 hr. incubation 1 Clarridge, Johnson, Pezzlo, and Weissfeld, ASM Cumitech 2B, November 1998. Dr.T.V.Rao MD 40
  41. 41. CUMITECH GUIDELINES FOR INTERPRETATION OF ROUTINE URINE CULTURES 1One isolate at >10 4:Full ID andSusceptibilityOne or two gram-negative isolates at >10 5and other isolates atleast 10X less: Full IDand Susceptibility ofgram-negative isolates Dr.T.V.Rao MD 41
  42. 42. CUMITECH GUIDELINES FOR INTERPRETATION OF ROUTINE URINE CULTURES 1 Other patterns of isolates at >10 4: Presumptive ID only Ignore mixed urethral flora at <10 4 1 Clarridge, Johnson, Pezzlo, and Weissfeld, ASM Cumitech 2B, November 1998. Dr.T.V.Rao MD 42
  43. 43. CUMITECH GUIDELINES FOR INTERPRETATION OF SPECIAL OR UNCOMMON URINE CULTURES 1 One or two isolates at >10 2 to 10 5: Full ID and Susceptibility is essential before confirmed as uncommon isolates Dr.T.V.Rao MD 43
  44. 44. ASM MANUAL GUIDELINES FOR URINE CULTURE RESULTS LIKELY TO BE SIGNIFICANT 1 Midstream, female with cystitis, >10 2 with positive urine leukocyte esterase Midstream, female with pyelonephritis, >10 5 with positive urine leukocyte esterase Midstream, asymptomatic, >10 5 with negative urine leukocyte esterase (usually) Midstream, male with UTI: >10 3 with leukocyte with urine leukocyte esterase positive Straight catheter: >10 2 with urine leukocyte esterase positive Indwelling catheter: >10 3 with urine leukocyte esterase positive or negative1 Manual of Clinical Microbiology, 8 th Edition, ASM, 2003 Dr.T.V.Rao MD 44
  45. 45. NMH GUIDELINES FOR INTERPRETATION OF URINE CULTURES 1 Urine leukocyte esterase positive One or two organisms at >10 3 : Full ID and Susceptibility One organism at >10 4 with others (2 or more) more ) at least 10X less: Full ID and susceptibility of predominant organism Report all group B β - hemolytic streptococci for women < 50 years1 Modified from ASM Cumitech, ASM Manual, and CDC MMWR 2002;51 (RR - 11):1- 22 (RR- 11):1 - Dr.T.V.Rao MD 45
  46. 46. NMH GUIDELINES FOR INTERPRETATION OF URINE CULTURES 1Urine leukocyte esterase negativeOne or two organisms at >10 5: Full andSusceptibilityOne4 gram-negative organism (pure culture) at>10 : Full ID and SusceptibilityYeast in pure culture: ID as Candida albicans or notC. albicansReport all group B β-hemolytic streptococci for women <50 years1 M od ified from ASM Cumitech, ASM M anual, and CDC M M WR 2002;51 (RR - 11):1 -22 Dr.T.V.Rao MD 46
  47. 47. ANTIBIOTIC SENSITIVIT Y TESTINGAll the isolatedbacteria identifiedas pathogenic to betested for AntibioticSensitivity/Resistancepattern by diskdiffusion methods Dr.T.V.Rao MD 47
  48. 48. ANTIBIOTIC SENSITIVITYAlways use a puregrowth of the isolate.Perform testing forantibiotic sensitivitywith inoculum whichis comparable todefined McFarlandstandards Dr.T.V.Rao MD 48
  49. 49. ANTIBIOTIC SENSITIVITY TESTINGHigh potency disksusedAntibiotics normallyactive in urineselectedAntibiotics found inhigher concentrationin urine preferredPrimary sensitivity testfrequently used Dr.T.V.Rao MD 49
  50. 50. REPORTING OF CONTAMINATED SPECIMENSMicroscopy helps todetect pus cells andepithelial cells, aninsignificant growth withfew pus cells can ignoredas contaminantsIn females even thepresence of leucocyteswith Squamous epithelialcells without a definedsignificant growth shouldbe ignored. Dr.T.V.Rao MD 50
  51. 51. REPORTING OF NEGATIVE URINE CULTURESNegative cultureresults showing nobacterial growth areavailable after 24hours. Positive resultsrequire 24-72 hours tocompleteidentification of thenumber and type ofbacteria found Dr.T.V.Rao MD 51
  52. 52. CLINICAL PROBLEMS MANIFESTING AS URINARY TRACT INFECTIONSAn infection of theGenitourinary tractwhere the MicrobesDonot grow on selectedmedia as inMycobacteriumtuberculosis Gonococcus Nutritionally exacting or anaerobic bacteriashould be considered. Dr.T.V.Rao MD 52
  53. 53. URINARY TRACT INFECTION ARE OVER DIAGNOSED Most over diagnosed condition, both by the clinicians and microbiologists. Makes the physicians to miss the existing condition. Unnecessary antibiotic prescription lead to drug resistance, loss of resources Dr.T.V.Rao MD 53
  54. 54. Created by Dr.T.V.Rao MD for ‘eLearning” resources forMicrobiologists in the Developing World Email doctortvrao@gmail.com Dr.T.V.Rao MD 54
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