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Practical of Urinary Tract Infection

Practical of Urinary Tract Infection. Department of Microbiology Medical Faculty University Of Indonesia. Important aspects of Microbiologic Examination of UTI: Specimen collection and handling - Specimen processing and culture - Interpretation of microbiology laboratory result.

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Practical of Urinary Tract Infection

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  1. Practical of Urinary Tract Infection Department of Microbiology Medical Faculty University Of Indonesia

  2. Important aspects of Microbiologic • Examination of UTI: • Specimen collection and handling • - Specimen processing and culture • - Interpretation of microbiology • laboratory result

  3. Microbiology of Community-Acquired Urinary Tract Infection Dysuria-Pyuria Sydrome in Females Other Community-Acquired Infection

  4. Microbiology of NosocomialAcquired Urinary Tract Infection in Children or Adult Catheter-Associated Short-Term (< 30 –d) Catheterization Catheter-Associated Long-Term (> 30 –d) Catheterization

  5. The urinary catheter Urine specimens for laboratory investigations can be collected from catheterized patients as shown (left). The second port is for putting fluids into the bladder (right). Urine from the drainage bag should not be tested because it may have been standing for several hours.

  6. Sampling Port Urine collection can be performed without needle Avoidance of needle stick injury

  7. TRANSPORT MEDIA Urine specimen is best collected Soon after a patient wakes when Organisms have had the opportunity to multiply over several hours. Urine container criteria : Sterile, dry, wide-necked, leak proof

  8. Laboratory examination of urine Quantitative (Colony counts) a urine sample is streaked on surface of Blood Agar plate ( for control) and medium Endo agar / Mc Conkey agarwith a special loop calibrated to deliver a known volume. + 1 + 2 + 3 + 4 Over night incubation Isolation of colonies,Biochemical tests, Drug susceptibility test, Over night incubation RESULT Urinary Tract infection Module’05 …..

  9. Area for colony count Blood Agar

  10. +1 Blood Agar ENDO Agar

  11. +2 Blood Agar ENDO Agar

  12. +3 Blood Agar ENDO Agar

  13. +4 Blood Agar ENDO Agar

  14. INTERPRETATION OF COLONY COUNTS Urinary Tract infection Module’05 …..

  15. BIOCHEMICAL TEST Carbohydrate fermentation Urinary Tract infection Module’05 …..

  16. Commercially identification The API 20Etm (produced by Biomerieux) system for bacteria such as coliform (ONPG : orthonitrophenyl- -D-galactopyramoside; VP : Voges-Proskauer)

  17. Three API 20E strips : • Immediately after inoculation • After 24 hours incubation • That in ( b) after addition of reagents to certain wells. • The organisms here is Escherichia coli. Here the first carbohydrate well (glucose) • is also used for the nitrate reduction test

  18. Antibiotic susceptibility tests • Disc Method • By using Disc which contain antibiotic • Diameter of Inhibition zone Bacterial growth show a response to antibiotic (NCCLS/CLSI standar) • Density standard : Mc Farland 0,5

  19. Disc susceptibility test Urinary Tract System Module’05 …..

  20. Gentamisin (CN) : 12 - 15 Chloramphenicol (C) : 12 - 18 Penicilin (P) : 28 - 29 R (Resisten) ; I (Intermediet) ; S (Sensitif) P P C C Staphylococcus aureus CN CN

  21. Tube method • One set of antibiotic dilution in liquid medium penipisan antibiotik dalam perbenihan cair, kemudian ditanami bakteri uji • Sensitivity are count by observation where the bacteria still growth in lowest antibiotic concentration (MIC)

  22. MIC : bakteri Escherichia coli = 2 g

  23. E-test strip

  24. The common organisms that can cause UTI And antibiotics to use in treatment

  25. Blood agar • An enriched medium, especially for culturing fastidious microorganism and observed the hemolytic reaction

  26. Endo C agar cat. No. 4044 Selective culture medium for detection and isolation Of fecal Escherichia coliandcoliformbacteria in Various material according to ENDO Sodium sulfide anf fuchsin inhibit the growth of Gram positive bacteria. E. coli and coliform bacteria metabolize lactose with the production of aldehyde and acid. The aldehyde liberates fuchsin from the fuchsin-sulfide compound, the fuchsin then colors the colonies red. E. coli will giving the permanent greenish metallic sheen (Fuchsin sheen)

  27. MacConkey Agar A differential medium for detection, isolation and enumeration of coliform and intestinal pathogen in water, dairy product and biological specimens. This agar is particularly recommended for the cultivation of pathogens which may be present in a variety of specimens such as urine, faeces and wound swabs. MacConkey agar should be used in parallel with other selective indicator media such as Desoxycholate Citrate Agar, Bismuth sulphite agar, Briliant Green Agar and BrilliantGreen bile broth, and non selective medium such as Blood Agar.

  28. NOVOBIOCIN TEST This test using for differentiated between Staphylococcus epidermidis and Staphylococcus saprophyticus from Urine specimens. A. Staphylococcus epidermidis (sensitive-Novobiocin), occasionally cause of urinary infection in hospital, sometimes associated with catheterization. B. Staphylococcus saprophyticus (resistant-Novobiocin), it causes cystitis in young women patients in general practice beside Escherichia coli Urinary Tract System Module’05 …..

  29. C.L.E.D Medium (Cystine-Lactose-Electrolyte Deficient) This medium recommended for urinary bacteriology, supporting the growth of all urinary pathogens. The present of important contaminants such as diphtheroids, lactobacilli and micrococci is also clearly elicited giving indicate and indication of the degree of contamination. This medium prevent the swarming phenomenon by having the low electrolyte content. Discrete colonies of motile and non motile bacteria may thus be urinary bacteriology where motile species, notably proteus spp. may be present a problem in identifying the species responsible to UTI.

  30. Proteus This species causes urinary infection, especially following catheterization or cystoscopy. Mostly Proteus strains produce a characteristic swarming growth over the surface of blood agar and several other culture media. Proteus culture have a distinctive smell. In laboratory C.L.E.D medium (electrolyte deficient) provide for plate culture of urinary organisms for prevents the swarming phenomenon Proteus spp. posses urease which produces ammonia in the urine. This raises the the pH and causes precipitation of phosphate crystals, leading to stone formation. Urinary Tract System Module’05 …..

  31. Pseudomonas Pseudomonas aeruginosa: This species causes urinary infection, usually following catheterization or associated with chronic urinary infections. This species is usually resistant to many antibiotics and is particularly associated with recurrent urinary tract infection in hospital patients. Urinary Tract System Module’05 …..

  32. Agar Endo

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