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    Diagnostic Techniques in Bacteriology

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    This document discusses several diagnostic techniques used in medical microbiology, including microscopy, culture-based techniques, serology, and molecular methods. Microscopy can be used to directly visualize microorganisms and includes techniques like simple staining, Gram staining, acid-fast staining, and immunofluorescence. Culture-based techniques aim to isolate and grow microbes. Serology detects antigens or antibodies. Molecular diagnostic methods allow detection of microbial nucleic acids.

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    Diagnostic Techniques in Bacteriology

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    355 views17 pages
    This document discusses several diagnostic techniques used in medical microbiology, including microscopy, culture-based techniques, serology, and molecular methods. Microscopy can be used to directly visualize microorganisms and includes techniques like simple staining, Gram staining, acid-fast staining, and immunofluorescence. Culture-based techniques aim to isolate and grow microbes. Serology detects antigens or antibodies. Molecular diagnostic methods allow detection of microbial nucleic acids.

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    This document discusses several diagnostic techniques used in medical microbiology, including microscopy, culture-based techniques, serology, and molecular methods. Microscopy can be used to directly visualize microorganisms and includes techniques like simple staining, Gram staining, acid-fast staining, and immunofluorescence. Culture-based techniques aim to isolate and grow microbes. Serology detects antigens or antibodies. Molecular diagnostic methods allow detection of microbial nucleic acids.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    355 views17 pages

    Diagnostic Techniques in Bacteriology

    Uploaded by

    api-19969058
    This document discusses several diagnostic techniques used in medical microbiology, including microscopy, culture-based techniques, serology, and molecular methods. Microscopy can be used to directly visualize microorganisms and includes techniques like simple staining, Gram staining, acid-fast staining, and immunofluorescence. Culture-based techniques aim to isolate and grow microbes. Serology detects antigens or antibodies. Molecular diagnostic methods allow detection of microbial nucleic acids.

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    Attribution Non-Commercial (BY-NC)
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    Download as ppt, pdf, or txt
    of 17

    Diagnostic techniques in

    medical microbiology

    Dr. Mejbah Uddin Ahmed


    Diagnosis of infectious diseases
     Infectious disease is diagnosed by:
     Clinically
     Radio logically
     Laboratory investigation
     microbiological

     hematological and other


    Diagnostic microbiology

    Patient

    Microbiology
    Laboratory

    Diagnosis
    Diagnostic microbiology

     Several procedures are involved in the


    diagnostic microbiology:
     Different types of microscopy.
     Cultural method.
     Serological method.
     Molecular method.
    Microscopy
     Microscopy: Is done for direct or indirect
    visualization of organism. Different types of
    microscopy are available:
     Light microscopy:
    -Unstained preparation.
    -Stined preparation.
     Immunoflourescence.
     Electronmicroscopy.
    Microscopy
    Simple stain: A single basic dye such as: methylene
    blue or basic fuchsin is used as simple stain.
    Negative stain: This technique is useful in the
    demonstration of bacterial capsule.
    Impregnation stain: Demonstration of spirochetes
    and bacterial flagella.
    Differential staining: They impart different colours
    to different bacteria (Gram’s stain and acid -fast
    stain.)
    Simple staining

     A single staining agent is used such as


    methylene blue, carbol fuchsin, crystal
    violet or safranin.
     It highlights the entire microorganism.
     Cellular shapes and basic structures are
    visible.
    Gram stain
     Developed by the Danish bacteriologist
    Christian Gram in 1884.
     Classifies bacteria into two groups based on
    differences in cell wall structure-
    Gram – positive & Gram- negative
     The gram stain is a very important early step
    in some medical situations may provide
    enough information to select an antibiotic.
    Steps of Gram Stain
    Ziehl - Neelsen
    1. Primary Stain by heated carbol fuchsin.
    2. Decolorize with 20% sulfuric acid and
    alcohol separately or with acid alcohol
    (which contains hydrochloric acid and
    ethanol).
    3. Wash the slide in water.
    4. Counterstain usually with methylene blue or
    malachite green.
    Immunofluorescence
     Direct:
     detection of antigen by use of
    fluorescence labeled antibodies
     used for diagnosis of viral infection,
    Chlamydia, legionella, Treponema
    pallidum, Giardia, Pneumocystis
     Indirect:
     detection of antibody.
    Electron-microscopy

    Important diagnostic technique for many


    viral infections
    Culture

     Used in bacterial, fungal, viral, parasitic


    diagnosis
     Mainstay of bacterial diagnosis
     Liquid media based culture

     Solid agar based culture

     Ensures organism available for


     susceptibility testing

     epidemiological studies
    Serology

     Depend on the detection of either


    antibody and/or antigen in patients
    serum.
     Antigen detection useful in acute
    diagnosis
     Antibody detection may be delayed
    Serology
     Types of tests:
     Precipitation.
     Agglutination.
     Complement fixation (CFT).
     Radioimmunoassay (RIA).
     Enzyme immunoassays (EIA)
     Immunochromatographic test (ICT).
     Enzymelinked Immunosorbant assay(ELISA)
    Molecular techniques
     Allows the detection of either RNA or
    DNA
     Western blotting - protein detection
     Southern blotting - DNA detection
     Polymerase chain reaction - DNA or
    RNA detection

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