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Overview of Diagnostic Microbiology and Immunology Lab Procedures

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40 diapositives · 620 Views · Aug 14, 2014

This chapter provides insights into diagnostic microbiology and immunology laboratory procedures, emphasizing specimen analysis, including urine tests using dipsticks and specific bacteriological media like Eosin Methylene Blue Agar. It covers essential tests such as glucose, protein, and antibiotic sensitivity assessments (Kirby-Bauer method). Additionally, it elaborates on biosafety levels in laboratories and provides details on disease progression in typhoid fever through antibody titers. The chapter also discusses various immunological techniques, including agglutination and enzyme immunoassays.

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  1. Chapter 31 Diagnostic Microbiology and Immunology

  2. Lab Diagnostic Time Lines

  3. The Swab Clicker Question:

  4. Here’s Some, Depending on Specimen Clicker Question

  5. Urine Analysis Dip Sticks Top: After reaction with metabolites in urine. Bottom: Before reaction with metabolites in urine Test time = 60 sec. Measures: glucose, bilirubin, ketones, sp. gravity, pH, protein, urobilinogen, nitrite, esterases. Top two right are false positives to show the color reaction.

  6. Eosin Methylene Blue Agar

  7. There are hundreds of Selective and Differential Bacteriological Media Text goes on describing particular samples: wounds, feces, genital swabs, etc. We will cover those when we get to specific diseases.

  8. Urethral Swab: Microscopy + Plating on Thayer-Martin Agar Oxidase Test Reagent poured onto the agar AFTER the colonies grew up.

  9. Gas Pack, Anaerobic Chamber

  10. Some Clinical Tests

  11. Metabolic Reactions The Classic API-20e Glucose Phenol Red with Durham tube. TSI: triple sugar iron agar. Citrate Test

  12. More Clinical Tests

  13. Testing Antibiotic Sensitivity Kirby-Bauer Zone of Inhibition E-test. MIC: Minimal Inhibitory Concentration

  14. Some Antibiotics – Interpretation of Size of the Zone of Inhibition

  15. Pseudomonas aeruginosaMICs Each numbered row – one antibiotic, highest conc at left, lowest at right. Clicker Question:

  16. E test Clicker Question:

  17. Working in a BSL-4 Lab The Gen Micro Labs in AHC2 are BSL-1, most our research labs are BSL-2. BSL-3 labs: to contain pathogens that could be infective as an aerosol. BSL-1: lab people wear lab coats, gloves..and usually does not involve pathogens. BSL-2: needs to contain pathogens in hoods, etc. Aerosol generating equipment must be in a biosafety cabinet.

  18. Disease Progression: Untreated Typhoid Fever Antibody Titers: Generally done by serial 1:2 dilutions from a blood sample. Highest dilution that brings about an Ab-Ag reaction. Clicker Question:

  19. Disease Progression: Untreated Typhoid Fever Antibody Titers: Generally done by serial 1:2 dilutions from a blood sample. Highest dilution that brings about an Ab-Ag reaction. Clicker Question

  20. Monoclonal Antibodies

  21. Precipitin Reaction

  22. Precipitin Rxns on a Glass Slide Clicker Question:

  23. Agglutination Reactions

  24. Latex Bead Agglutination for S. aureus

  25. Fluorescent Antibodies Clostridium septicumstained with FITC-Ab, C. chauvoelstained with Rhodamine-Ab

  26. Direct and Sandwich (Indirect)

  27. B and T Cells… Yellow-Green cells are CD8 Cells Red-orange cells are CD-4 Cells

  28. Cell Sorting Clicker Question:

  29. Enzyme Immuno-Assays

  30. EIA Kits for Small Clinics

  31. Immunoblots: Detecting Blotted Proteins (after electrophoresis)

  32. Nucleic Acid Diagnostics

  33. PCR Detecting Mycobacteium tuberculosis Lanes 1 + 10 Molecular Weight Standards Lanes 2-12 from different TB patients sputum Lanes 13-14 from non-TB patients with lung symptoms. Lane 15 – from known Mycobacterium tuberculosis DNA.