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Urinary Tract Infection Microbiology Practical Class

This practical class aims to teach students about specimen collection and processing, microbiological and biochemical analysis of urine specimens, culture and identification of organisms, antibiotics susceptibility testing, and interpretation of results. Students will also learn about clinically important etiological organisms associated with UTIs, their identification, and susceptibility testing.

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Urinary Tract Infection Microbiology Practical Class

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  1. URINARYTRACT INFECTION MicrobiologyPracticalClass RenalSystemBlock FirstYear Dr.FawziaAl-Otaibi

  2. Objectives Itisexpectedthatbytheendofthispracticalclass,studentsshould beableto: 1. 2. 3. Knowtheimportantstepsinspecimencollectionandtransportto thelab. HowtoprocessurineSpecimensinthelab. –Urinemicrobiologicalandbiochemicalanalysis. –Organismscultureandidentification. –Antibioticsusceptibilitytesting. –Resultsinteroperation. KnowtheclinicallyimportantetiologicalOrganismsassociated withUTI,theiridentificationandsusceptibilitytesting.

  3. SPECIMEN COLLECTION

  4. TYPEOFSPECIMENS Midstreamurine(MSU) Adhesivebag SuprapubicAspiration Cathetersample

  5. TYPEOFSPECIMENS

  6. SterileUrineContainer Dipslides OnesideisCLEDmedia,the othercanbeMacConkey(MAC) agarorbloodagar.

  7. SPECIMEN PROCESSING

  8. Specimenprocessing: 1.Urineanalysis •Microbiological •Biochemical 2.Cultureandidentification(ID) 3.AntimicrobialSusceptibility testing 4.Resultsinterpretation

  9. 1.UrineAnalysis Biochemical Microbiological Macroscopic Microscopic

  10. BiochemicalUrine Examination(Dipstick) •leukocyteesterase •Nitratetest •PH •Glucose •Bilirubin •Protein

  11. TestingforUTI Midstreamcleancatchwithdipstickanalysis Nitrite+→forgram-negativebacteriawhichcanconvertnitrate tonitrite(sensitivity92-100%,lowspecificity); falsenegativewithbacteriathatdonotreducenitrate gram-positivebacteria excessdietaryVitaminC Leukocyteesterase+→indicatespresenceofwhitebloodcells, (sensitivity75-95%,specificity94-98%)– Dipstickresultsmaybeaffectedbymedications/dyes,iepyridium, nitrofurantoin,metronidazole,bilirubin,methyleneblue,VitaminB complex

  12. PhysicalUrineExamination Macroscopic •Color •Odor •Turbidity

  13. MicrobiologicalUrine Examination Microscopic: •Cell-counting(WBC,RBC) •Parasite •Casts (Ovum,Trichomonas,yeast)

  14. MicroscopicUrine Examination(WETMOUNT)

  15. 2.CULTUREANDID

  16. CultureandIdentification Culturemedia Urineinoculationandreadingofculture Identificationofculturedorganisms

  17. CultureMediaRoutinely UsedforUrineCulture

  18. UrineInoculation Quantitative(Colonycounts) +1 +2 +3 +4 aurinesampleisstreakedonsurfaceof BloodAgarplateandCLEDagar/Mc Conkeyagarwithaspecialloop calibratedtodeliveraknownvolume. Overnightincubation Isolationofcolonies, Biochemicaltests, Drugsusceptibilitytest, Overnightincubation RESULT

  19. Smi-QantitativeCultureof UrineSample

  20. IDofculturedorganisms Biochemicaltests. Typeofhemolysis Serologicaltests

  21. CLINICALLYIMPORTANT MICROORGANISMS CAUSINGUTI

  22. EtiologicalAgentsofUTI Bacteria Fungi Parasites

  23. BACTERIA Gram-negativebacilli Enterobacteriace • •Non-Enterobacteriace Gram-positivecocci Staphylococci Coagulase-positive(Staph.aureus) Coagulasenegative(Staph.saprophyticus,Staph.   Epidermidis) Streptococci Streptococcus(groupB) Enterococci  

  24. GramNegativeBacilli Enterobacteriacae Non-Enterobacteriacae

  25. Enterobacteriacae •Escherichiacoli •Klebseilla •Proteus •Enterobacter •Citrobacter

  26. E.coli

  27. Klebseillaspp

  28. Proteusspp

  29. Non-Enterobacteriacae Pseudomonasspp

  30. GramPositiveCocci Staphylococci Coagulase-positive(Staph.aureus) Coagulasenegative(Staph.saprophyticus) Coagulasenegative(Staph.epidermidis)    Streptococcus Enterococci (groupB)

  31. Staph.aureus

  32. Staph.epidermidis

  33. Staph.saprophyticus

  34. Strept.agalactiae (groupB)

  35. Enterococci

  36. FUNGI

  37. Candidaalbicans

  38. PARASITESCAUSINGUTI Schistosomahaematobium

  39. FUNGICAUSINGUTI Candidaalbicans

  40. Schistosomahaematobium (urine;eggs115-170x45-65micrometers) (primates)

  41. 3.Antimicrobial SusceptibilityTesting (AST)

  42. MethodsofAST Diskdiffusiontest Etest

  43. TheAntibioticSensitivityTestingMethod

  44. DiskDiffusionMethod

  45. ETEST •E-TEST •Isawell-establishedmethodfor antimicrobialresistancetesting inmicrobiologylaboratories. •Consistsofapredefined gradientofantibiotic concentrationsonaplasticstrip •Usedtodeterminethe MinimumInhibitory Concentration(MIC)of –Antibiotics –Antifungalagents –Antimycobacterialagents

  46. Enterobacteriacae AntibioticSusceptibilityTest •FirstlineUTItreatment: Ampicillin TMP/SMX Ciprofloxacin Gentamicin Amikacin Nitrofunatoin – – – – – –

  47. BacterialResistancetoUTIAntibiotics Nitrofurantoin –E.coli<5% –Otheruropathogens15-20% –Notactiveagainst •Proteus, •someEnterobacter •Klebsiella TMP-SMX(Bactrim) 10-22%variesgeographically Fluoroquinolones(Cipro) therapyforacuteuncomplicatedcystitis Ampicillin –30%ofE.coliresistant

  48. Emergingresistantto AmpicillinandTMP/SMX Interpretation R S S R S S S S Antibiotic Ampicillin Cephalexin Ciprofloxacin TMP/SMX Nitrofurantoin Gentamicin Ceftriaxone Imipenem Organism Source MIC ≥32 ≤4 ≤4 ≥2 ≤16 ≤8 ≤1 ≤1 Ecoli urine

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