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<title>Disk diffusion test</title>

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Disk diffusion test

</h1>

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</style><div role="note" class="hatnote navigation-not-searchable">Not to be confused with the <a href="Kleihauer%E2%80%93Betke_test" title="Kleihauer–Betke test">Kleihauer–Betke test</a>, which is also often called a "KB test".</div>

The disk diffusion test (also known as the agar diffusion test, Kirby–Bauer test, disc-diffusion antibiotic susceptibility test, disc-diffusion antibiotic sensitivity test and KB test) is a <a href="Microbiological_culture" title="Microbiological culture">culture</a>-based <a href="Microbiology" title="Microbiology">microbiology</a> assay used in <a href="Diagnostic" class="mw-redirect" title="Diagnostic">diagnostic</a> and <a href="Drug_discovery" title="Drug discovery">drug discovery</a> laboratories. In diagnostic labs, the assay is used to determine the susceptibility of bacteria isolated from a patient's infection to clinically approved antibiotics. This allows physicians to prescribe the most appropriate antibiotic treatment.<a href="#cite_note-EUCAST2021a-1">[1]</a><a href="#cite_note-pmid1214010-2">[2]</a><a href="#cite_note-pmid13669774-4">[4]</a><a href="#cite_note-pmid5325707-5">[5]</a> In drug discovery labs, especially <a href="Bioprospecting" title="Bioprospecting">bioprospecting</a> labs, the assay is used to screen biological material (e.g. plant extracts, bacterial fermentation broths) and drug candidates for antibacterial activity. When bioprospecting, the assay can be performed with paired strains of bacteria to achieve dereplication and provisionally identify antibacterial <a href="Mechanism_of_action" title="Mechanism of action">mechanism of action</a>.<a href="#cite_note-pmid32529587-6">[6]</a><a href="#cite_note-pmid21819327-7">[7]</a>

In diagnostic laboratories, the test is performed by inoculating the surface of an agar plate with bacteria isolated from a patient's infection. Antibiotic-containing paper disks are then applied to the agar and the plate is incubated. If an antibiotic <a href="Bacteriostatic_agent" title="Bacteriostatic agent">stops the bacteria from growing</a> or <a href="Bacteriocidal" class="mw-redirect" title="Bacteriocidal">kills the bacteria</a>, there will be an area around the disk where the bacteria have not grown enough to be visible. This is called a zone of inhibition. The susceptibility of the bacterial isolate to each antibiotic can then be semi-quantified by comparing the size of these zones of inhibition to databases of information on known antibiotic-susceptible, moderately susceptible and resistant bacteria. In this way, it is possible to identify the most appropriate antibiotic for treating a patient's infection.<a href="#cite_note-EUCAST2021a-1">[1]</a><a href="#cite_note-pmid1214010-2">[2]</a> Although the disk diffusion test cannot be used to differentiate bacteriostatic and bactericidal activity, it is less cumbersome than other susceptibility test methods such as <a href="Minimum_inhibitory_concentration#Broth_dilution_assay" title="Minimum inhibitory concentration">broth dilution</a>.<a href="#cite_note-pmid13669774-4">[4]</a>

In drug discovery labs, the disk diffusion test is performed slightly differently than in diagnostic labs. In this setting, it is not the bacterial strain that must be characterized, but a test extract (e.g. a plant or microbial extract). The agar plate is therefore inoculated with a bacterial strain of known phenotype (often an <a href="ATCC_(company)" title="ATCC (company)">ATCC</a> or <a href="National_Collection_of_Type_Cultures" title="National Collection of Type Cultures">NCTC</a> strain), and disks containing the test extract are applied to the surface (see <a href="#Quality_control">below</a>).<a href="#cite_note-pmid32529587-6">[6]</a> Zone of inhibition sizes cannot be used as a semi-quantitative measure of antibacterial potency because different extracts contain molecules with different diffusion characteristics (different <a href="Molecular_size" class="mw-redirect" title="Molecular size">molecular sizes</a>, <a href="Hydrophile" title="Hydrophile">hydrophilicities</a> etc.). Zone of inhibition sizes can be used for the purpose of dereplication though. This is achieved by testing each extract against paired strains of bacteria (e.g. streptomycin-susceptible and -resistant strains to identify streptomycin-containing extracts). Paired strains (e.g. wild type and <a href="Biological_target" title="Biological target">target</a> overexpressing strains) can also be used to identify antibacterial mechanism of action.<a href="#cite_note-pmid32529587-6">[6]</a><a href="#cite_note-pmid21819327-7">[7]</a>

<div class="mw-heading mw-heading2"><h2 id="History">History</h2></div>

Agar diffusion was first used by <a href="Martinus_Beijerinck" title="Martinus Beijerinck">Martinus Beijerinck</a> in 1889 to study the effect of <a href="Auxins" class="mw-redirect" title="Auxins">auxins</a> on bacterial growth. However, the method has been developed, refined and standardized by many scientists and scientific organizations over the years including George F. Reddish, <a href="Norman_Heatley" title="Norman Heatley">Norman Heatley</a>, James G. Vincent,<a href="#cite_note-pmid11420332-8">[8]</a> Alfred W. Bauer, William M.M. Kirby, <a href="John_C._Sherris" class="mw-redirect" title="John C. Sherris">John C. Sherris</a>,<a href="#cite_note-pmid13669774-4">[4]</a><a href="#cite_note-pmid5325707-5">[5]</a> Hans Martin Ericsson, the <a href="World_Health_Organization" title="World Health Organization">World Health Organization</a>, the <a href="Clinical_and_Laboratory_Standards_Institute" title="Clinical and Laboratory Standards Institute">Clinical and Laboratory Standards Institute</a>, the Swedish Reference Group for Antibiotics, the <a href="Deutsches_Institut_f%C3%BCr_Normung" title="Deutsches Institut für Normung">Deutsches Institut für Normung</a>, the <a href="British_Society_for_Antimicrobial_Chemotherapy" title="British Society for Antimicrobial Chemotherapy">British Society for Antimicrobial Chemotherapy</a> and others.<a href="#cite_note-pmid11420332-8">[8]</a>

<div class="mw-heading mw-heading2"><h2 id="Principle">Principle</h2></div>

A pure bacterial culture is suspended in saline, its turbidity is standardized, and it is swabbed uniformly across an agar plate. An antibiotic- or extract-impregnated filter paper disk is then placed on the surface of the agar. The disk constituent(s) diffuse from the filter paper into the agar. The concentration of these constituents will be highest next to the disk and will decrease as the distance from the disk increases. If the antibiotic or extract is effective against bacteria at a certain concentration, no colonies will grow where the concentration in the agar is greater than or equal to the effective concentration. This is the zone of inhibition. In general, larger zones of inhibition correlate with lower <a href="Minimum_inhibitory_concentration" title="Minimum inhibitory concentration">minimum inhibitory concentrations</a> (MICs) of antibiotic or extract for that bacterial strain.<a href="#cite_note-EUCAST2021a-1">[1]</a> An exception to this is when molecules of the antibiotic or extract are large or hydrophobic because these diffuse through the agar slowly.<a href="#cite_note-pmid32529587-6">[6]</a>

<div class="mw-heading mw-heading2"><h2 id="Standard_method">Standard method</h2></div>

<div class="mw-heading mw-heading3"><h3 id="Agar_plate_and_inoculum_preparation">Agar plate and inoculum preparation</h3></div>

All aspects of the Kirby–Bauer procedure are standardized to ensure consistent and accurate results. Because of this, a laboratory must adhere to these standards. The media used in Kirby–Bauer testing must be <a href="Mueller%E2%80%93Hinton_agar" title="Mueller–Hinton agar">Mueller–Hinton agar</a> at only 4 mm deep, poured into either 100 mm or 150 mm Petri dishes. The <a href="PH" title="PH">pH</a> level of the agar must be between 7.2 and 7.4. Bacterial inoculum is prepared by diluting a broth culture to match a 0.5 <a href="McFarland_standards" title="McFarland standards">McFarland turbidity standard</a>, which is equivalent to approximately 150 million <a href="Cell_(biology)" title="Cell (biology)">cells</a> per mL.<a href="#cite_note-EUCAST2021a-1">[1]</a>

<div class="mw-heading mw-heading3"><h3 id="Inoculation_and_incubation">Inoculation and incubation</h3></div>

Using <a href="Aseptic_technique" class="mw-redirect" title="Aseptic technique">aseptic technique</a>, <a href="Growth_medium" title="Growth medium">broth</a> <a href="Cell_culture" title="Cell culture">culture</a> of a specific organism is collected with a <a href="Sterilization_(microbiology)" title="Sterilization (microbiology)">sterile</a> <a href="Cotton_swab" title="Cotton swab">swab</a>. In the case of <a href="Gram_negative_bacteria" class="mw-redirect" title="Gram negative bacteria">Gram negative bacteria</a>, excess liquid is removed from the swab by gently pressing or rotating it against the inside of the tube. The swab is then streaked across a Mueller–Hinton agar plate to form a bacterial lawn. To obtain uniform growth, the agar plate is streaked with the swab in one direction, rotated 120° and streaked again, rotated another 120° and streaked again. Using an antibiotic disk dispenser, disks containing specific antibiotics are then applied to the plate. This must be done within 15 minutes of inoculation. Flame-sterilized forceps are used to gently press each disk onto the agar and ensure it is attached. Plates are then <a href="Incubator_(culture)" title="Incubator (culture)">incubated</a> overnight, usually at a temperature of 35 °C. Plates must be incubated within 15 minutes of applying antibiotic disks.<a href="#cite_note-EUCAST2021a-1">[1]</a>

<div class="mw-heading mw-heading3"><h3 id="Test_media_modifications">Test media modifications</h3></div>

Certain bacteria require addition of 5% solution of mechanically defibrinated horse blood and <a href="Nicotinamide_adenine_dinucleotide" title="Nicotinamide adenine dinucleotide">β-NAD</a> (MH-F agar).<a href="#cite_note-EUCAST2021a-1">[1]</a> The following table shows media requirements of commonly tested microorganisms:

<caption>Disk diffusion media requirements<a href="#cite_note-EUCAST2021a-1">[1]</a>

</caption>

<th>Standard MH agar

</th>

<th>MH-F agar

</th></tr>

<tr>

<td>

<ul><li>Enterobacteriales</li>

<li>Pseudomonas spp.</li>

<li>Stenotrophomonas maltophilia</li>

<li>Acinetobacter spp.</li>

<li>Staphylococcus spp.</li>

<li>Enterococcus spp.</li>

<li>Aeromonas spp.</li>

<li>Achromobacter xylosoxidans</li>

<li>Vibrio spp.</li>

<li>Bacillus spp.</li>

<li>Burkholderia pseudomallei</li></ul>

</td>

<td>

<ul><li>Streptococcus groups A, B, C, G</li>

<li>Streptococcus pneumoniae</li>

<li>Streptococcus viridans group</li>

<li>Haemophilus influenzae</li>

<li>Moraxella catarrhalis</li>

<li>Listeria monocytogenes</li>

<li>Pasteurella multocida</li>

<li>Campylobacter jejuni and C. coli</li>

<li>Corynebacterium</li>

<li>Aerococcus sanguinicola and A. urinae</li>

<li>Kingella kingae</li>

<li>Brucella melitensis</li></ul>

</td></tr></tbody></table>

<div class="mw-heading mw-heading3"><h3 id="Quality_control">Quality control</h3></div>

To ensure veracity of test results, <a href="Quality_control" title="Quality control">quality control</a> methods must be used. In order to monitor the performance of the test, special bacterial strains are used as positive or negative control. When efficacy of β-lactamase is tested, special strains that exhibit β-lactam resistance are used. Additionally, specific media are used to test certain antibiotics. For example, when testing <a href="Co-trimoxazole" class="mw-redirect" title="Co-trimoxazole">co-trimoxazole</a> susceptibility, media with excess <a href="Thymine" title="Thymine">thymine</a> and <a href="Thymidine" title="Thymidine">thymidine</a> are recommended.<a href="#cite_note-EUCAST2021a-1">[1]</a> The following table lists commonly used quality control strains in the disk diffusion method:

<caption>Quality control strains (as of January 2025)<a href="#cite_note-EUCAST2021a-1">[1]</a>

</caption>

<th rowspan="2">Bacterium

</th>

<th colspan="6">Strain

</th>

<th rowspan="2">Description

</th>

<th rowspan="2">Antibiotics tested

</th></tr>

<tr>

<th>ATCC

</th>

</th>

</th>

</th>

</th>

</th></tr>

<tr>

<td rowspan="3">E. coli

</td>

</td>

<td>12241

</td>

<td>76.24

</td>

<td>1103

</td>

<td>17620

</td>

<td>434

</td>

<td>susceptible (wild type)

</td>

<td><a href="Neomycin" title="Neomycin">neomycin</a>, <a href="Colistin" title="Colistin">colistin</a>, <a href="Kanamycin" class="mw-redirect" title="Kanamycin">kanamycin</a>, <a href="Cephalexin" class="mw-redirect" title="Cephalexin">cephalexin</a>, <a href="Gentamicin" title="Gentamicin">gentamicin</a>, <a href="Cefamandole" title="Cefamandole">cefamandole</a>, <a href="Cefalotin" title="Cefalotin">cephalotin</a>, <a href="Tetracycline" title="Tetracycline">tetracycline</a>, <a href="Cephaloglycin" class="mw-redirect" title="Cephaloglycin">cephaloglycin</a>, <a href="Cephaloridine" title="Cephaloridine">cephaloridine</a>, <a href="Nalidixic_acid" title="Nalidixic acid">nalidixic acid</a>, <a href="Chloramphenicol" title="Chloramphenicol">chloramphenicol</a><a href="#cite_note-:0-14">[14]</a>

</td></tr>

<tr>

</td>

<td>11954

</td>

<td>102181

</td>

<td>5923

</td>

<td>30600

</td>

<td>943

</td>

<td>produce <a href="TEM-1" class="mw-redirect" title="TEM-1">TEM-1</a> β-lactamase, resistant to ampicillin (used to check β-lactamase component of β-lactam combination disks)

</td>

<td>

</td></tr>

<tr>

<td>-

</td>

</td>

<td>-

</td>

<td>-

</td>

<td>-

</td>

<td>-

</td>

<td>produces <a href="Beta-lactamase#CTX-M_beta-lactamases_(class_A)" title="Beta-lactamase">CTX-M-15</a> and <a href="Beta-lactamase#OXA_beta-lactamases_(class_D)" title="Beta-lactamase">OXA-1</a> (used to check β-lactamase component of β-lactam combination disks)

</td>

<td><a href="Cefotaxime" title="Cefotaxime">cefotaxime</a><a href="#cite_note-:1-16">[16]</a>

</td></tr>

<tr>

<td rowspan="2">Klebsiella pneumoniae

</td>

</td>

<td>13368

</td>

<td>-

</td>

<td>-

</td>

<td>45421

</td>

<td>7787

</td>

<td>produces <a href="Beta-lactamase#SHV_beta-lactamases_(class_A)" title="Beta-lactamase">SHV-18</a> (an extended-spectrum β-lactamase, used to check β-lactamase component of β-lactam combination disks)

</td>

<td>

</td></tr>

<tr>

</td>

<td>-

</td>

<td>-

</td>

<td>-

</td>

<td>-

</td>

<td>-

</td>

<td>produces <a href="Beta-lactamase#KPC_(K._pneumoniae_carbapenemase)_(class_A)" title="Beta-lactamase">KPC-3</a>, SHV-11, TEM-1 (used to check β-lactamase component of β-lactam combination disks)

</td>

<td>novel β-lactam/β-lactamase inhibitor combinations (e.g., <a href="Meropenem/vaborbactam" title="Meropenem/vaborbactam">meropenem/vaborbactam</a>)<a href="#cite_note-:2-19">[18]</a>

</td></tr>

<tr>

<td>Pseudomonas aeruginosa

</td>

</td>

<td>12903

</td>

<td>76.110

</td>

<td>1117

</td>

<td>17619

</td>

<td>108

</td>

<td>susceptible (wild type)

</td>

<td>

</td></tr>

<tr>

<td rowspan="2">Staphylococcus aureus

</td>

</td>

<td>12973

</td>

<td>103429

</td>

<td>2569

</td>

<td>15915

</td>

<td>794

</td>

<td>produces β-lactamases (weak)

</td>

<td>

</td></tr>

<tr>

<td>-

</td>

</td>

<td>-

</td>

<td>-

</td>

<td>67181

</td>

<td>

</td>

<td><a href="Methicillin-resistant_Staphylococcus_aureus" title="Methicillin-resistant Staphylococcus aureus">MRSA</a> (<a href="MecA" title="MecA">mecA</a> plasmid-positive)

</td>

<td><a href="Methicillin" title="Methicillin">methicillin</a> and other antibiotics affected by MRSA strains<a href="#cite_note-:3-22">[21]</a>

</td></tr>

<tr>

<td rowspan="2">Enterococcus faecalis

</td>

</td>

<td>12697

</td>

<td>103214

</td>

<td>2570

</td>

<td>9997

</td>

<td>795

</td>

<td>susceptible (wild type)

</td>

<td>

</td></tr>

<tr>

</td>

<td>13379

</td>

<td>104676

</td>

<td>12956

</td>

<td>34289

</td>

<td>-

</td>

<td><a href="HLAR" class="mw-redirect" title="HLAR">HLAR</a> (aminoglycoside-modyfing enzyme), resistant to vancomycin (<a href="Vancomycin_resistance" class="mw-redirect" title="Vancomycin resistance">vanB</a> plasmid-positive)

</td>

<td>gentamicin, <a href="Streptomycin" title="Streptomycin">streptomycin</a><a href="#cite_note-:4-24">[23]</a>

</td></tr>

<tr>

<td>Streptococcus pneumoniae

</td>

</td>

<td>12977

</td>

<td>104340

</td>

<td>11967

</td>

<td>33638

</td>

<td>-

</td>

<td>resistant to <a href="Benzylpenicillin" title="Benzylpenicillin">benzylpenicillin</a>

</td>

<td>

</td></tr>

<tr>

<td rowspan="2">Haemophilus influenzae

</td>

</td>

<td>12975

</td>

<td>103570

</td>

<td>11970

</td>

<td>29539

</td>

<td>-

</td>

<td>susceptible (wild type)

</td>

<td>

</td></tr>

<tr>

</td>

<td>12699

</td>

<td>104604

</td>

<td>9999

</td>

<td>26214

</td>

<td>-

</td>

<td>reduced suscibility to β-lactams (exhibits modified <a href="Penicillin_binding_proteins" class="mw-redirect" title="Penicillin binding proteins">penicillin binding proteins</a>)

</td>

<td>

</td></tr>

<tr>

<td>Campylobacter jejuni

</td>

</td>

<td>11351

</td>

<td>70.2T

</td>

<td>4688

</td>

<td>11284

</td>

<td>-

</td>

<td>susceptible (wild type), requires microaerobic environment and higher incubation temperature (41±1°C)

</td>

<td>

</td></tr></tbody></table>

<div class="mw-heading mw-heading2"><h2 id="Alternate_methods">Alternate methods</h2></div>

Several variations of the disk diffusion method have been developed including the Oxford penicillin cup and <a href="Etest" title="Etest">Etest</a> methods used in hospital diagnostic laboratories,<a href="#cite_note-pmid18339637-29">[28]</a><a href="#cite_note-pmid31596673-30">[29]</a> and the well diffusion, cylinder diffusion and bioautography methods used in drug discovery and development laboratories.<a href="#cite_note-pmid32529587-6">[6]</a><a href="#cite_note-pmid33148158-31">[30]</a>

<div class="mw-heading mw-heading3"><h3 id="Oxford_penicillin_cup_method">Oxford penicillin cup method</h3></div>

Disks containing increasing antibiotic concentrations are placed on a seeded bacterial lawn on the agar surface and plates are incubated. Zone sizes are measured from the edge of the disk to the end of the clear zone. Interpretation is more complicated in mixed susceptibility populations. These are plotted as linear dimensions or squares of distances as a function of the natural logarithm of antibiotic concentration in the disks. The MIC is determined from the zero intercept of a linear regression fit through the data.<a href="#cite_note-agdif-32">[31]</a> The intercept itself is the logarithm of the MIC. The slope of the regression line is related to the diffusion coefficient of that particular antibiotic in the agar.<a href="#cite_note-pmid18339637-29">[28]</a>

<div class="mw-heading mw-heading3"><h3 id="EUCAST_Rapid_Antibiotic_Susceptibility_Test_(RAST)">EUCAST Rapid Antibiotic Susceptibility Test (RAST)</h3></div>

The RAST method serves as a fast means of ascertaining antibiotic susceptibility and was created as a modification of the classic disk diffusion test. It allows to shorten the time of incubation to 16-20 hours. Test scores are read after 4, 6, 8 and 16-20 hours. Compared to the standard method, RAST does not give distinct zones of inhibition within such a short timespan (all bacteria except for S. pneumoniae have a chance of being possible to read after 6 hours higher than 90%). As for quality control strains, they are diluted 1:1 000 000 and defibrinated horse or sheep blood is added. Special RAST breakpoint tables should be used when interpreting the results due to method calibration differences.<a href="#cite_note-33">[32]</a>

Validated quality control strains include: E. coli ATCC 25922, P. aeruginosa ATCC 27853, S. aureus ATCC 29213, E. faecalis ATCC 29212, S. pneumoniae ATCC 49619.<a href="#cite_note-34">[33]</a>

<div class="mw-heading mw-heading4"><h4 id="Screening_for_antibiotic_resistance_mechanisms_in_RAST">Screening for antibiotic resistance mechanisms in RAST</h4></div>

RAST allows for rapid determination of possible antibiotic resistance in tested cultures. It allows to check for ESBL and/or carbapenemase producing E. coli and K. pneumoniae, using cefotaxime/ceftazidime (after 4 hours) and meropenem (after 6 hours) respectively. However, these results are not quantitative and should be used only for screening in routine medical tests.<a href="#cite_note-35">[34]</a><a href="#cite_note-36">[35]</a> In clinical trials, RAST method led to significant improvements in predicting efficacy of antibiotic therapy.<a href="#cite_note-37">[36]</a>

<div class="mw-heading mw-heading3"><h3 id="Disk_pre-diffusion_method">Disk pre-diffusion method</h3></div>

Disks containing antibiotics are placed on an uninoculated Mueller-Hinton agar plate and incubated for 2 hours. Then, they are removed and the bacterial suspension previously prepared using broth microdilution is applied and another disk with a different antibiotic is placed precisely in the same place as the previous one. After incubation for 16-20 hours results are correlated with the first antibiotic's MIC values. An example of the pre-diffusion method is testing in vitro efficacy of ceftazidime/avibactam (primary disks) in terms of aztreonam (secondary disks).<a href="#cite_note-38">[37]</a>

<div class="mw-heading mw-heading3"><h3 id="Antifungal_drug_testing">Antifungal drug testing</h3></div>

Disk diffusion method can be used to test susceptibility to antifungals.<a href="#cite_note-39">[38]</a><a href="#cite_note-40">[39]</a>

<div class="mw-heading mw-heading3"><h3 id="Bioautography">Bioautography</h3></div>

Comparing to classical disk-based methods, bioautography utilises <a href="Thin-layer_chromatography" title="Thin-layer chromatography">thin-layer chromatography</a> to separate constituents of the tested mixture. Then, the TLC plate can be either placed on the inoculated agar and be allowed to diffuse into it (contact bioautography) or be covered with microbe-containing broth (direct bioautography). Then, the sample is incubated and zones of inhibitions are measured.<a href="#cite_note-:6-42">[41]</a><a href="#cite_note-43">[42]</a><a href="#cite_note-:5-41">[40]</a> Alternatively, the TLC plate can be covered with molten agar in the agar overlay bioautography.<a href="#cite_note-44">[43]</a>

To visualise zones of inhibitions in direct bioautography, reagents that detect <a href="Dehydrogenase" title="Dehydrogenase">dehydrogenase</a> activity are used (e.g., <a href="Tetrazolium_salts" class="mw-redirect" title="Tetrazolium salts">tetrazolium salts</a>, which are converted by microbial dehydrogenases into chromogenic <a href="Formazan" title="Formazan">formazans</a>).<a href="#cite_note-:6-42">[41]</a>

<div class="mw-heading mw-heading2"><h2 id="Other_images">Other images</h2></div>

<div class="gallerytext">Agar diffusion was first used in 1889 by <a href="Martinus_Beijerinck" title="Martinus Beijerinck">Martinus Beijerinck</a>.<a href="#cite_note-pmid11420332-8">[8]</a></div>

</li>

<div class="gallerytext">A close-up look at the results of an agar diffusion test</div>

</li>

<div class="gallerytext">An antibiogram of <a href="Serratia_marcescens" title="Serratia marcescens">Serratia marcescens</a>. Each disk is labelled with the antibiotic it contains (e.g. AMC30, 30 μg <a href="Amoxicillin/clavulanic_acid" title="Amoxicillin/clavulanic acid">amoxicillin/clavulanic acid</a>)</div>

</li>

<div class="gallerytext">Automated culture plate reader to measure inhibition zone diameters</div>

</li>

</ul>

<ul><li><a href="Antibiotic_sensitivity_testing" title="Antibiotic sensitivity testing">Antibiotic sensitivity testing</a></li>

<li><a href="Double-disk_diffusion_test" title="Double-disk diffusion test">Double-disk diffusion test</a></li>

<li><a href="Etest" title="Etest">Etest</a></li></ul>

<div class="mw-heading mw-heading2"><h2 id="References">References</h2></div>

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<li id="cite_note-38"><a href="#cite_ref-38">^</a> <cite id="CITEREFLimade_LimaRochaSampaio2022" class="citation journal cs1">Lima, Keila de Oliveira; de Lima, Aline Valério; Rocha, Darlan Augusto da Costa; Sampaio, Suely Carlos Ferreira; Cappellano, Paola; Sampaio, Jorge Luiz Mello (March 2022). <a rel="nofollow" class="external text" href="https://doi.org/10.1016%2Fj.jgar.2021.12.009">"A simple disk pre-diffusion test to predict in vitro aztreonam/avibactam activity against NDM-producing Klebsiella pneumoniae complex"</a>. Journal of Global Antimicrobial Resistance. 28: 49–52. <a href="Doi_(identifier)" class="mw-redirect" title="Doi (identifier)">doi</a>:<a rel="nofollow" class="external text" href="https://doi.org/10.1016%2Fj.jgar.2021.12.009">10.1016/j.jgar.2021.12.009</a>. <a href="PMID_(identifier)" class="mw-redirect" title="PMID (identifier)">PMID</a> <a rel="nofollow" class="external text" href="https://pubmed.ncbi.nlm.nih.gov/34936924">34936924</a>.</cite>

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<li id="cite_note-39"><a href="#cite_ref-39">^</a> <cite id="CITEREFOzkutukErgonMetinYucesoy2008" class="citation journal cs1">Ozkutuk, A.; Ergon, C.; Metin, D.Y.; Yucesoy, M.; Polat, S.H. (February 2008). <a rel="nofollow" class="external text" href="http://www.tandfonline.com/doi/full/10.1179/joc.2008.20.1.87">"Comparison of Disk Diffusion, E-Test and Broth Microdilution Test in Determination of Susceptibility of Aspergillus Species to Amphotericin B, Itraconazole and Voriconazole"</a>. Journal of Chemotherapy. 20 (1): 87–92. <a href="Doi_(identifier)" class="mw-redirect" title="Doi (identifier)">doi</a>:<a rel="nofollow" class="external text" href="https://doi.org/10.1179%2Fjoc.2008.20.1.87">10.1179/joc.2008.20.1.87</a>. <a href="ISSN_(identifier)" class="mw-redirect" title="ISSN (identifier)">ISSN</a> <a rel="nofollow" class="external text" href="https://search.worldcat.org/issn/1120-009X">1120-009X</a>. <a href="PMID_(identifier)" class="mw-redirect" title="PMID (identifier)">PMID</a> <a rel="nofollow" class="external text" href="https://pubmed.ncbi.nlm.nih.gov/18343749">18343749</a>.</cite>

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<li id="cite_note-40"><a href="#cite_ref-40">^</a> <cite id="CITEREFKronvallKarlsson2001" class="citation journal cs1">Kronvall, Göran; Karlsson, Inga (April 2001). <a rel="nofollow" class="external text" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC87949">"Fluconazole and Voriconazole Multidisk Testing of Candida Species for Disk Test Calibration and MIC Estimation"</a>. Journal of Clinical Microbiology. 39 (4): 1422–1428. <a href="Doi_(identifier)" class="mw-redirect" title="Doi (identifier)">doi</a>:<a rel="nofollow" class="external text" href="https://doi.org/10.1128%2FJCM.39.4.1422-1428.2001">10.1128/JCM.39.4.1422-1428.2001</a>. <a href="ISSN_(identifier)" class="mw-redirect" title="ISSN (identifier)">ISSN</a> <a rel="nofollow" class="external text" href="https://search.worldcat.org/issn/0095-1137">0095-1137</a>. <a href="PMC_(identifier)" class="mw-redirect" title="PMC (identifier)">PMC</a> <a rel="nofollow" class="external text" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC87949">87949</a>. <a href="PMID_(identifier)" class="mw-redirect" title="PMID (identifier)">PMID</a> <a rel="nofollow" class="external text" href="https://pubmed.ncbi.nlm.nih.gov/11283066">11283066</a>.</cite>

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<li id="cite_note-:5-41">^ <a href="#cite_ref-:5_41-0">a</a> <a href="#cite_ref-:5_41-1">b</a> <cite id="CITEREFConde-MartínezAcosta-GonzálezDíazTello2017" class="citation journal cs1">Conde-Martínez, Natalia; Acosta-González, Alejandro; Díaz, Luis E.; Tello, Edisson (December 2017). <a rel="nofollow" class="external text" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5721385">"Use of a mixed culture strategy to isolate halophilic bacteria with antibacterial and cytotoxic activity from the Manaure solar saltern in Colombia"</a>. BMC Microbiology. 17 (1): 230. <a href="Doi_(identifier)" class="mw-redirect" title="Doi (identifier)">doi</a>:<a rel="nofollow" class="external text" href="https://doi.org/10.1186%2Fs12866-017-1136-x">10.1186/s12866-017-1136-x</a>. <a href="ISSN_(identifier)" class="mw-redirect" title="ISSN (identifier)">ISSN</a> <a rel="nofollow" class="external text" href="https://search.worldcat.org/issn/1471-2180">1471-2180</a>. <a href="PMC_(identifier)" class="mw-redirect" title="PMC (identifier)">PMC</a> <a rel="nofollow" class="external text" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5721385">5721385</a>. <a href="PMID_(identifier)" class="mw-redirect" title="PMID (identifier)">PMID</a> <a rel="nofollow" class="external text" href="https://pubmed.ncbi.nlm.nih.gov/29216824">29216824</a>.</cite>

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<li id="cite_note-:6-42">^ <a href="#cite_ref-:6_42-0">a</a> <a href="#cite_ref-:6_42-1">b</a> <cite id="CITEREFChomaGrzelak2011" class="citation journal cs1">Choma, Irena M.; Grzelak, Edyta M. (May 2011). <a rel="nofollow" class="external text" href="https://linkinghub.elsevier.com/retrieve/pii/S0021967310017632">"Bioautography detection in thin-layer chromatography"</a>. Journal of Chromatography A. 1218 (19): 2684–2691. <a href="Doi_(identifier)" class="mw-redirect" title="Doi (identifier)">doi</a>:<a rel="nofollow" class="external text" href="https://doi.org/10.1016%2Fj.chroma.2010.12.069">10.1016/j.chroma.2010.12.069</a>. <a href="PMID_(identifier)" class="mw-redirect" title="PMID (identifier)">PMID</a> <a rel="nofollow" class="external text" href="https://pubmed.ncbi.nlm.nih.gov/21232747">21232747</a>.</cite>

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<li id="cite_note-43"><a href="#cite_ref-43">^</a> <cite id="CITEREFWangZhangWangWang2021" class="citation journal cs1">Wang, Meng; Zhang, Yirong; Wang, Ruijie; Wang, Zhibin; Yang, Bingyou; Kuang, Haixue (2021-07-31). <a rel="nofollow" class="external text" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8347725">"An Evolving Technology That Integrates Classical Methods with Continuous Technological Developments: Thin-Layer Chromatography Bioautography"</a>. Molecules. 26 (15): 4647. <a href="Doi_(identifier)" class="mw-redirect" title="Doi (identifier)">doi</a>:<a rel="nofollow" class="external text" href="https://doi.org/10.3390%2Fmolecules26154647">10.3390/molecules26154647</a>. <a href="ISSN_(identifier)" class="mw-redirect" title="ISSN (identifier)">ISSN</a> <a rel="nofollow" class="external text" href="https://search.worldcat.org/issn/1420-3049">1420-3049</a>. <a href="PMC_(identifier)" class="mw-redirect" title="PMC (identifier)">PMC</a> <a rel="nofollow" class="external text" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8347725">8347725</a>. <a href="PMID_(identifier)" class="mw-redirect" title="PMID (identifier)">PMID</a> <a rel="nofollow" class="external text" href="https://pubmed.ncbi.nlm.nih.gov/34361800">34361800</a>.</cite>

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<li id="cite_note-44"><a href="#cite_ref-44">^</a> <cite id="CITEREFNuthanRakshithMarulasiddaswamyRao2020" class="citation journal cs1">Nuthan, Bettadapura Rameshgowda; Rakshith, Devaraju; Marulasiddaswamy, Kuppuru Mallikarjunaiah; Rao, H C Yashavantha; Ramesha, Kolathur Puttamadaiah; Mohana, Nagabhushana Chandra; Siddappa, Shiva; Darshan, Doreraj; Kumara, Kigga Kaadappa Sampath; Satish, Sreedharamurthy (2020-08-21). <a rel="nofollow" class="external text" href="https://academic.oup.com/chromsci/article/58/8/737/5882178">"Application of Optimized and Validated Agar Overlay TLC–Bioautography Assay for Detecting the Antimicrobial Metabolites of Pharmaceutical Interest"</a>. Journal of Chromatographic Science. 58 (8): 737–746. <a href="Doi_(identifier)" class="mw-redirect" title="Doi (identifier)">doi</a>:<a rel="nofollow" class="external text" href="https://doi.org/10.1093%2Fchromsci%2Fbmaa045">10.1093/chromsci/bmaa045</a>. <a href="ISSN_(identifier)" class="mw-redirect" title="ISSN (identifier)">ISSN</a> <a rel="nofollow" class="external text" href="https://search.worldcat.org/issn/0021-9665">0021-9665</a>. <a href="PMID_(identifier)" class="mw-redirect" title="PMID (identifier)">PMID</a> <a rel="nofollow" class="external text" href="https://pubmed.ncbi.nlm.nih.gov/32766714">32766714</a>.</cite>

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</ol></div>

<div class="mw-heading mw-heading2"><h2 id="Notes">Notes</h2></div>

<li id="cite_note-18"><a href="#cite_ref-18">^</a> Klebsiella quasipneumoniae Brisse et al

</li>

</ol></div></div>

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</style><div id="Techniques_in_clinical_microbiology192" style="font-size:114%;margin:0 4em">Techniques in <a href="Clinical_microbiology" class="mw-redirect" title="Clinical microbiology">clinical microbiology</a></div></th></tr><tr><th scope="row" class="navbox-group" style="width:1%"><a href="Isolation_(microbiology)" title="Isolation (microbiology)">Isolation</a> and <a href="Culture_(microbiology)" class="mw-redirect" title="Culture (microbiology)">culture</a></th><td class="navbox-list-with-group navbox-list navbox-odd hlist" style="width:100%;padding:0"><div style="padding:0 0.25em"></div><table class="nowraplinks navbox-subgroup" style="border-spacing:0"><tbody><tr><th scope="row" class="navbox-group" style="width:1%">Isolation techniques</th><td class="navbox-list-with-group navbox-list navbox-odd" style="width:100%;padding:0"><div style="padding:0 0.25em">

<ul><li><a href="Asepsis" title="Asepsis">Asepsis</a></li>

<li><a href="Streaking_(microbiology)" title="Streaking (microbiology)">Streak plate</a></li>

<li><a href="Selective_media" class="mw-redirect" title="Selective media">Selective media</a></li></ul>

</div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%">Cultures by body site</th><td class="navbox-list-with-group navbox-list navbox-even" style="width:100%;padding:0"><div style="padding:0 0.25em">

<ul><li><a href="Blood_culture" title="Blood culture">Blood culture</a></li>

<li>Genital culture</li>

<li><a href="Sputum_culture" title="Sputum culture">Sputum culture</a></li>

<li><a href="Throat_culture" title="Throat culture">Throat culture</a></li>

<li><a href="Urine_culture" class="mw-redirect" title="Urine culture">Urine culture</a></li>

<li><a href="Wound_culture" class="mw-redirect" title="Wound culture">Wound culture</a></li></ul>

</div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%">Cultures by organism</th><td class="navbox-list-with-group navbox-list navbox-odd" style="width:100%;padding:0"><div style="padding:0 0.25em">

<ul><li><a href="Bacterial_culture" class="mw-redirect" title="Bacterial culture">Bacterial culture</a></li>

<li>Fungal culture</li>

<li><a href="Viral_culture" title="Viral culture">Viral culture</a></li></ul>

</div></td></tr></tbody></table><div></div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%"><a href="Microbiological_identification" class="mw-redirect" title="Microbiological identification">Identification</a> and testing</th><td class="navbox-list-with-group navbox-list navbox-odd hlist" style="width:100%;padding:0"><div style="padding:0 0.25em"></div><table class="nowraplinks navbox-subgroup" style="border-spacing:0"><tbody><tr><th scope="row" class="navbox-group" style="width:1%">Manual testing: basic techniques</th><td class="navbox-list-with-group navbox-list navbox-even" style="width:100%;padding:0"><div style="padding:0 0.25em">

<ul><li><a href="Colonial_morphology" title="Colonial morphology">Colonial morphology</a>

<ul><li><a href="Hemolysis_(microbiology)" title="Hemolysis (microbiology)">Hemolysis</a></li></ul></li>

<li><a href="Staining_(biology)" class="mw-redirect" title="Staining (biology)">Staining</a>

<ul><li><a href="Gram_stain" title="Gram stain">Gram stain</a></li>

<li><a href="Acid-fast_stain" class="mw-redirect" title="Acid-fast stain">Acid-fast stain</a></li>

<li><a href="Giemsa_stain" title="Giemsa stain">Giemsa stain</a></li>

<li><a href="India_ink_stain" class="mw-redirect" title="India ink stain">India ink stain</a></li>

<li><a href="Ziehl%E2%80%93Neelsen_stain" title="Ziehl–Neelsen stain">Ziehl–Neelsen stain</a></li></ul></li>

<li>Rapid tests

<ul><li><a href="Oxidase_test" title="Oxidase test">Oxidase</a></li>

<li><a href="Catalase_test" class="mw-redirect" title="Catalase test">Catalase</a></li>

<li><a href="Indole_test" title="Indole test">Indole</a></li>

</div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%">Manual testing: biochemical and immunologic tests</th><td class="navbox-list-with-group navbox-list navbox-odd" style="width:100%;padding:0"><div style="padding:0 0.25em">

<li>Amino acid decarboxylase test</li>

<li><a href="Bile_solubility_test" class="mw-redirect" title="Bile solubility test">Bile solubility test</a></li>

<li><a href="Citrate_test" title="Citrate test">Citrate test</a></li>

<li><a href="Coagulase_test" class="mw-redirect" title="Coagulase test">Coagulase test</a></li>

<li><a href="Diagnostic_microbiology#DNA_hydrolysis" title="Diagnostic microbiology">DNAse test</a></li>

<li><a href="IMViC" title="IMViC">IMViC</a></li>

<li><a href="Methyl_red_test" class="mw-redirect" title="Methyl red test">Methyl red test</a></li>

<li><a href="Diagnostic_microbiology#Nitrite_test" title="Diagnostic microbiology">Nitrite test</a></li>

<li><a href="Oxidative/fermentation_glucose_test" title="Oxidative/fermentation glucose test">Oxidative/fermentation glucose test</a></li>

<li><a href="Phenylalanine_deaminase_test" class="mw-redirect" title="Phenylalanine deaminase test">Phenylalanine deaminase test</a></li>

<li><a href="Diagnostic_microbiology#Reverse_CAMP_test" title="Diagnostic microbiology">Reverse CAMP test</a></li>

<li><a href="Diagnostic_microbiology#6.5%_salt_broth" title="Diagnostic microbiology">Salt tolerance test</a></li>

<li><a href="Sulfide_indole_motility_test" class="mw-redirect" title="Sulfide indole motility test">Sulfide indole motility test</a></li>

<li><a href="Triple_sugar_iron_test" class="mw-redirect" title="Triple sugar iron test">Triple sugar iron test</a></li>

<li><a href="Urease#As_diagnostic_test" title="Urease">Urease test</a>

<ul><li><a href="Rapid_urease_test" title="Rapid urease test">rapid</a></li></ul></li>

<li><a href="Voges%E2%80%93Proskauer_test" title="Voges–Proskauer test">Voges–Proskauer test</a></li>

<li>X and V factor test</li>

<li>Bacitracin susceptibility test</li>

<li><a href="Optochin_susceptibility_test" class="mw-redirect" title="Optochin susceptibility test">Optochin susceptibility test</a></li>

<li><a href="Novobiocin_susceptibility_test" class="mw-redirect" title="Novobiocin susceptibility test">Novobiocin susceptibility test</a></li>

<li><a href="Lancefield_grouping" title="Lancefield grouping">Lancefield grouping</a></li>

</div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%">Automated and <a href="Point-of-care_testing" title="Point-of-care testing">point-of-care testing</a></th><td class="navbox-list-with-group navbox-list navbox-even" style="width:100%;padding:0"><div style="padding:0 0.25em">

<ul><li><a href="Analytical_profile_index" title="Analytical profile index">Analytical profile index</a></li>

<li><a href="MALDI-TOF" class="mw-redirect" title="MALDI-TOF">MALDI-TOF</a></li>

<li><a href="Polymerase_chain_reaction#Infectious_disease_applications" title="Polymerase chain reaction">Polymerase chain reaction</a></li>

<li><a href="VITEK" title="VITEK">VITEK</a></li>

<li><a href="Rapid_strep_test" title="Rapid strep test">Rapid strep test</a></li>

<li><a href="Monospot_test" class="mw-redirect" title="Monospot test">Monospot test</a></li></ul>

</div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%"><a href="Antibiotic_sensitivity" class="mw-redirect" title="Antibiotic sensitivity">Antibiotic susceptibility testing</a></th><td class="navbox-list-with-group navbox-list navbox-odd" style="width:100%;padding:0"><div style="padding:0 0.25em">

<ul><li>Beta-lactamase test</li>

<li><a href="Etest" title="Etest">Etest</a></li>

<li><a href="McFarland_standards" title="McFarland standards">McFarland standards</a></li>

<li><a href="Minimum_inhibitory_concentration" title="Minimum inhibitory concentration">Minimum inhibitory concentration</a></li></ul>

</div></td></tr></tbody></table><div></div></td></tr><tr><th scope="row" class="navbox-group" style="width:1%">Equipment</th><td class="navbox-list-with-group navbox-list navbox-even hlist" style="width:100%;padding:0"><div style="padding:0 0.25em">

<ul><li><a href="Agar_plate" title="Agar plate">Agar plate</a>

<ul><li><a href="Growth_medium" title="Growth medium">Growth medium</a></li></ul></li>

<li><a href="McIntosh_and_Filde's_anaerobic_jar" class="mw-redirect" title="McIntosh and Filde's anaerobic jar">Anaerobic jar</a>

<li><a href="Durham_tube" title="Durham tube">Durham tube</a></li>

<li><a href="Biosafety_cabinet" title="Biosafety cabinet">Biosafety cabinet</a></li>

<li><a href="Incubator_(culture)" title="Incubator (culture)">Incubator</a></li>

<li><a href="Inoculation_loop" title="Inoculation loop">Inoculation loop</a></li>

<li><a href="Inoculation_needle" title="Inoculation needle">Inoculation needle</a></li></ul>

</div></td></tr></tbody></table></div></div><div><div class="zim-footer">

This article is issued from <a class="external text" title="Last edited on 2025-07-04" href="https://en.wikipedia.org/wiki/?title=Disk_diffusion_test&oldid=1298691366">Wikipedia</a>. The text is available under <a class="external text" href="https://creativecommons.org/licenses/by-sa/4.0/deed.en">Creative Commons Attribution-Share Alike 4.0</a> unless otherwise noted. Additional terms may apply for the media files.

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