Cefoxitin CX 30 mcg
Intended Use
Cefoxitin CX 30 mcg discs are used for antimicrobial susceptibility testing of bacterial cultures as per Kirby-Bauer
Method.
Principle
Antimicrobial Susceptibility Testing (AST) is a laboratory procedure performed by laboratory technician to identify,
which antimicrobial regimen is specifically effective for individual patients. The introduction of various antimicrobials
for treating variety of infections showed the necessity of performing antimicrobial susceptibility testing as a routine
procedure in all microbiology laboratories. antibiotics are generally defined as the substances produced by the
microorganism such as Penicillium, which has the ability to kill or inhibit the growth of other microorganisms, mainly
bacteria. Antimicrobial Susceptibility Tests (ASTs) basically measures the ability of an antibiotic or other
antimicrobial agent to inhibit the In vitro microbial growth.
The basic principle of the antibiotic susceptibility testing has been used in microbiology laboratories over 80 years.
Till the 1950s, laboratories were lacking in the methodologies and equipment’s for the accurate determination of
In vitro responses of organisms to antimicrobial agents. Bauer et al., began the development of standardized
methods for antimicrobial susceptibility testing, using disc diffusion system. Antimicrobial Susceptibility Tests are
either quantitative or qualitative.
Clinical laboratories currently employ several methods depending on the laboratory test menu that they provide.
These approaches include the disk diffusion and Minimum Inhibitory Concentration (MIC) methods. Disc diffusion
test is a qualitative test method. The National Committee for Clinical Laboratory Standards (NCCLS), now known
as Clinical Laboratory Standards Institute (CLSI) has published comprehensive documents regarding the disc
diffusion systems. The agar disc diffusion test is the most convenient and widely used method for routine
Antimicrobial Susceptibility Testing. In subsequent and current practice, antimicrobial impregnated paper discs are
applied onto the agar surface.
Various regulatory agencies and standards-writing organizations, published standardized reference procedures
based on the Kirby-Bauer method. Standardized reference procedures for the disc systems were published by
WHO and FDA and are periodically updated by the CLSI (formerly NCCLS) for any antimicrobial testing, Quality
control or clinical testing. However, few precautions are to be maintained while handling of the Sensitivity discs,
the latest CLSI documents should be consulted for current recommendations.
Susceptibility Test Procedure
Prepare plates with Mueller Hinton Agar (201130650100 / 201130650500, 203130830250 / 203130830100,
203090070015 / 203090070030 & Ready Prepared Plates – 205131790100 / 205131790020) for use in the
Kirby-Bauer Method for rapidly growing aerobic organisms. For fastidious organisms such as Streptococci, the
agar (201130650100 / 201130650500) is supplemented with 5% sterile, defibrinated blood. For Haemophilus
spp., Haemophilus Test Medium (for all agents when testing H. influenzae or H. parainfluenzae) or MH-F agar
(MHA with 5% mechanically defibrinated horse blood and 20 µg/mL NAD) (for selected agents when testing H.
influenzae); for N. gonorrhoeae, GC Agar Base (201070010500) with 1% defined growth supplement and for
Streptococcus pneumoniae, MHA with 5% sheep blood or MH-F agar (MHA with mechanically defibrinated
horse blood and 20 µg/mL NAD) are recommended respectively. The medium in the plates should be sterile
and have a depth of about 4 mm.
Bacterial Inoculum Preparation
1. Gram staining is done before starting susceptibility testing to confirm the culture purity and to determine
appropriate battery of tests.
2. Select 3-4 colonies from subcultured slant or plate and transfer them into 5 mL of sterile 0.85% saline,
vortex the suspension and adjust the turbidity to yield a uniform suspension matching 0.5 McFarland
standard (O.D at 630 nm range: 0.08-0.130).
Note: The direct colony suspension method is preferred for Staphylococcus spp., S. pneumoniae and
other Streptococci, Haemophilus spp. and N. gonorrhoeae. OR
3-5 well-isolated colonies should be selected and transferred to 4-5 mL of a suitable broth using an
inoculating needle.
3. Incubate the broth (usually 2-6 hours) at 35°C-37°C until it achieves or exceeds the turbidity of the 0.5
McFarland's barium sulphate standard. This results in a suspension containing approximately 1-2 X 108
cfu/mL.
Note: Overnight broth cultures should not be used as inoculum.
4. Adjust the turbidity to the barium sulphate standard. For the diluents use sterile broth or sterile saline. The
turbidity of the standard and the test inoculums should be compared by holding both tubes in front of a
white background with finely divided lines or by use of a photometric device.
5. Within 15 minutes of adjusting the turbidity of the inoculum, immerse a sterile cotton swab into the properly
diluted inoculum and rotate it firmly several times against the upper inside wall of the tube to express
excess fluid.
6. Inoculate the entire agar surface of the plate three times, rotating the plate 60° between streaking to
obtain even inoculation. Swab the rim of the agar bed too.
7. The lid may be left ajar for 3-5 minutes and the plate held at room temperature for not more than 15
minutes to allow the surface moisture to be absorbed before applying the antibiotic discs.
8. Apply discs by means of an antimicrobial disc dispenser, aseptically, at least 24 mm apart. Preferably,
deposit Penicillin and Cephalosporin discs not more than 10 mm from the edge of the Petri dish, and their
centers at least 30 mm apart. Avoid placing such discs adjacent to one another. Tap the discs with some
sterile needle or forceps after placing them on the agar for complete contact with the medium surface.
9. Within 15 minutes of applying the discs, invert the plates and incubate at 35°C-37°C. With non-fastidious
organisms the plates should not be incubated under an increased concentration of carbon dioxide.
Note: For Staphylococcus spp., testing at temperatures above 35°C may not detect Methicillin-resistant
Staphylococci (MRS); for N. gonorrhoeae, incubate at 35°C-37°C [do not exceed 37°C]. Haemophilus
spp., N. gonorrhoeae, S. pneumoniae and other Streptococci should be incubated in an atmosphere
enriched with 5% CO2.
10. Examine plates after 18-24 hours of incubation. 20-24 hours for N. gonorrhoeae, S. pneumoniae and
other Streptococci). A full 24 hours of incubation is recommended to detect methicillin/ oxacillin/
vancomycin-resistant Staphylococci in Staphylococcus spp., cefoxitin in Staphylococcus spp. (except
from S. aureus, S. lugdunensis, S. pseudintermedius and S. schleiferi), and Enterococcus spp. in
vancomycin resistance. The diameters of the zones of complete inhibition are measured, as determined
by gross visual inspection.
Zones are measured to the nearest whole millimeter. For further details in measuring zones of inhibition, consult
the reference. If only isolated colonies grow, the inoculum is too light and the test should be repeated. Zones
around discs containing different drugs are not comparable for the purpose of comparing activity of drugs. See the
Zone Diameter Interpretive Chart, which gives expected values from testing common aerobes.
Interpretation
Antimicrobial Agent Interpretative criteria for Resistant Intermediate Sensitive
(mm / ≤) (mm) (mm / ≥)
Cefoxitin 30 mcg Enterobacterales 14 15-17 18
Staphylococcus spp., except: 24 - 25
S. aureus
S. lugdunensis
S. epidermidis
S. pseudintermedius
S. schleiferi
Neisseria gonorrhoeae 23 24-27 28
Quality Control
Appearance: Filter paper disc of 6 mm diameter with printed ‘’CX 30 mcg’’ on each side of the disc.
Cultural response: Average diameter of zone of inhibition observed on Mueller Hinton Agar for 18-24 hours
incubation at 35°C-37°C for standard cultures.
Organism (ATCC) Standard Diameter of zone of
inhibition in mm
Escherichia coli (25922) 23-29
Staphylococcus aureus subsp. aureus 23-29
(25923)
Neisseria gonorrhoeae (49226) 33-41
Storage and Shelf-life:
Discs in routine use should be stored at 2°C-8°C. Longer term storage should be at -20°C-8°C.
References:
1. Bauer, Kirby, Sherris and Turck, 1966, Am. J. Clin. Path., 45: 493
2. Performance standards of Antimicrobial Disc Susceptibility Tests, CLSI Vol. 34, Feb 2024.
3. Data on file: Microxpress®, A Division of Tulip Diagnostics (P) Ltd.
Product Presentation:
Cat No. Product description Pack Size
206030510250 Antimicrobial Susceptibility Disc 5 Carts (5 x 50 disc)
206030510500 Antimicrobial Susceptibility Disc 5 Vials (5 x 100 disc)
206030510100 Antimicrobial Susceptibility Disc Single Vial (1 x 100 Disc)
