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    Agar Diluation

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    Maryam Adel
    The agar dilution method is a quantitative antibiotic susceptibility testing technique where serial dilutions of an antibiotic are incorporated into agar plates. A standardized bacterial suspension is inoculated onto the plates and incubated overnight. The minimum inhibitory concentration is determined as the lowest concentration of antibiotic that prevents visible bacterial growth. This method allows testing of multiple bacteria simultaneously on a single plate and determination of precise MIC values, making it well-suited for large-scale surveillance programs. However, it is time-consuming and labor-intensive, so agar dilution is generally reserved for reference laboratories.

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    Agar Diluation

    Uploaded by

    Maryam Adel
    171 views10 pages
    The agar dilution method is a quantitative antibiotic susceptibility testing technique where serial dilutions of an antibiotic are incorporated into agar plates. A standardized bacterial suspension is inoculated onto the plates and incubated overnight. The minimum inhibitory concentration is determined as the lowest concentration of antibiotic that prevents visible bacterial growth. This method allows testing of multiple bacteria simultaneously on a single plate and determination of precise MIC values, making it well-suited for large-scale surveillance programs. However, it is time-consuming and labor-intensive, so agar dilution is generally reserved for reference laboratories.

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    baily and scott diagnostic microbiology agar dilution

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    agar diluation

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    The agar dilution method is a quantitative antibiotic susceptibility testing technique where serial dilutions of an antibiotic are incorporated into agar plates. A standardized bacterial suspension is inoculated onto the plates and incubated overnight. The minimum inhibitory concentration is determined as the lowest concentration of antibiotic that prevents visible bacterial growth. This method allows testing of multiple bacteria simultaneously on a single plate and determination of precise MIC values, making it well-suited for large-scale surveillance programs. However, it is time-consuming and labor-intensive, so agar dilution is generally reserved for reference laboratories.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    171 views10 pages

    Agar Diluation

    Uploaded by

    Maryam Adel
    The agar dilution method is a quantitative antibiotic susceptibility testing technique where serial dilutions of an antibiotic are incorporated into agar plates. A standardized bacterial suspension is inoculated onto the plates and incubated overnight. The minimum inhibitory concentration is determined as the lowest concentration of antibiotic that prevents visible bacterial growth. This method allows testing of multiple bacteria simultaneously on a single plate and determination of precise MIC values, making it well-suited for large-scale surveillance programs. However, it is time-consuming and labor-intensive, so agar dilution is generally reserved for reference laboratories.

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    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    of 10

    Agar Dilution method

    • Agar dilution susceptibility testing is the solid equivalent of

    broth dilution susceptibility testing

    • Agar dilution is a quantitative susceptibility testing method

    because MIC values can be obtained using the method

    • This method is specially used if a single antibiotic is to be tested

    against a number of bacteria. Up to 30 different bacteria can be

    tested by this method at a time.


    • Standard media; Mueller-Hinton agar (MHA)
    • Antibiotic dilation preparation ; In this method, two-fold
    serial dilutions of an antibiotic
    • the range of antibiotic dilution needed Its based on the
    MIC range for organism,its different for each antibiotic
    • Each doubling dilution of an antimicrobial agent is
    incorporated in single agar plate.
    • a standardized bacterial suspension adjusted to 0.5
    McFarland
    • bacterial suspensions were inoculated on the MHA using
    a Cathra replicator with 1 mm pins, as recommended by
    the CLSI

     One control plate is inoculated without antibiotics.


    Temperature and Duration

     Most organisms should be incubated at 35°C to 37°C.


     The duration of incubation depends on the species.
    • For rapidly growing species such as the Enterobacteriaceae, the glucose-
    nonfermenting gram-negative bacilli, Enterococcus species, and Staphylococcus
    species, overnight incubation for a minimum of 16 hours is needed.
    • Longer incubation is recommended for fastidious species (18 to 20 hours plus),
    for Staphylococcus species when testing against antistaphylococcal penicillins (24
    hours), for Campylobacter species (24 to 48 hours depending on the standard and
    incubation temperature),
    • for Helicobacter pylori (3 days), and for anaerobes (42 to 48 hours).
    Result and interpretation

     The MIC is taken as the first concentration at which no growth occurs


     The organisms are reported sensitive, intermediate, or resistant by
    comparing the test MIC values with that given in CLSI guidelines.
    The key difference between agar diffusion and
    disc diffusion method
    is that in agar  diffusion method, the inoculum is filled into a hole or a
    well created on the agar medium while in agar disc diffusion method, a
    filter paper disc containing the antimicrobial solution is placed on the
    agar surface.

    7
    Advantages
    • The advantages of agar dilution include the ability to simultaneously
    test the susceptibility of a number of bacteria in one plate
    • the ability to test susceptibility of fastidious organisms since the agar
    with supplements is able to adequately support the bacteria growth.
    • Microbial contamination easily detected as compared with broth method
    • Moreover, as mentioned above, the test results yield MIC values for
    testing bacteria.
    • It is therefore well suited to the rapid evaluation of new compounds or for
    large-scale centralized surveillance programs
    8
    Disadvantages

    However, agar dilution is not commonly used in most microbiology


    laboratories due to the time consuming and labor- intensive

    • Agar dilution testing generally is not performed in


    routine clinical laboratories but can be ideal for regional
    reference laboratories or research laboratories that
    must test large numbers of isolates.

    9
    THANK
    YOU

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