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DISC DIFFUSION Method SHIMAA

The document describes the disk diffusion method for antibiotic susceptibility testing. This method involves preparing a bacterial suspension, streaking bacterial cultures on agar plates, applying antibiotic disks to the plates, incubating the plates overnight, and measuring any zones of inhibition around the disks to determine antibiotic susceptibility. Key steps include preparing a 0.5 McFarland standard inoculum, ensuring dried plates before applying disks spaced 1.5-2 cm apart, incubating plates overnight at 30-35°C, and measuring zones of inhibition in millimeters.

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Maryam Adel
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99 views25 pages

DISC DIFFUSION Method SHIMAA

The document describes the disk diffusion method for antibiotic susceptibility testing. This method involves preparing a bacterial suspension, streaking bacterial cultures on agar plates, applying antibiotic disks to the plates, incubating the plates overnight, and measuring any zones of inhibition around the disks to determine antibiotic susceptibility. Key steps include preparing a 0.5 McFarland standard inoculum, ensuring dried plates before applying disks spaced 1.5-2 cm apart, incubating plates overnight at 30-35°C, and measuring zones of inhibition in millimeters.

Uploaded by

Maryam Adel
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
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DIFFUSION METHOD

DISC DIFFUSION
(KIRBY_BAUER)
PRIMARY GOALS

ANNUAL
REVENU
E
GROWTH
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PRESENTATION TITLE
INTRODUCTION:
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DISK DIFFUSION: INOCULUM


PREPARATION
• A bacterial suspension with turbidity
equivalent to 0.5 MacFarland is prepared
in saline or nutrient broth.
• Soak a sterile cotton swab in the bacterial
suspension.
• Remove excess fluid by pressing the swab
against the tube wall

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PRESENTATION TITLE

Disk Diffusion: plate streaking

• Streak the swab evenly over the surface of the


Müller-Hinton medium in three directions to
ensure even distribution of bacteria.
• With petri dish lid in place, allow 3- 5 minutes
(no longer than 15 minutes) for agar surface to
dry
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PRESENTATION TITLE

DISK DIFFUSION: APPLYING


DISKS
• Using sterile thin forceps, place
antibiotic discs to the surface of
the agar.
• Antibiotic discs should be 1.5-2
cm spaced otherwise inhibition
zones will overlap.
• Within 30 minutes of applying
the discs, the plates are inverted
and incubated overnight at 30-35°
C.
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PRESENTATION TITLE U S E TR A N S M I TTE D LI G H T , RATH E R T H A N R EF L ECT ED L I G H T,
W H EN MEA SU RI N G ZO N ES

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M EA SU RI N G T H E ZO N E O F I N H I BI TI O N BY R U L ER O R M ET RI C I N MM I N
D I A MET ER

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• Do not use disks beyond their expiration date.


• Do not store disks in a frost-free freezer.
• Use FDA cleared products.
• Use disks with the content specified in CLSI standards.
• Do not relocate a disk once it has touched the agar
surface

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Click icon to add picture

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Measure the point at which you can see an obvious demarcation


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between growth and no growth. Measure the obvious zone.
Ignore the swarm even if it covers the zone
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• Zones with
trimethoprimsulfamethoxazole
(and also sulfonamides and
trimethoprim alone) may be
difficult to read because this
agent may not inhibit bacteria
from multiplying until the
bacteria have gone through
several generations of growth.
• You may see a light haze of
growth within the zone.
Measure the zone at the point
where there is an 80% reduction
in growth
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ADVANTAGE AND DISADVANTAGES
PRESENTATION TITLE

• Convenient and user-


friendly method

• It is generally not
accepted to test slow
growing bacteria that
require extended
incubation such as
mycobacteria and
anaerobes.

• Lack of interpretive
24 criteria for many organism
THANK YOU

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