1. Antibiogram realisation on a bacterial strain

2. Materials avalaible Swab Agar with bacteria Antibiotics disks Mueller Hinton culture medium Deposit plan Sterile physiological water tube

3. 1.Preparation of inoculum

4. 1.1. With a sterile inoculation loop, take a bacterial colony on agar Petri dish. WARNING! The sample is realized in asepsis around the Bunsen burner.

5. 1.2. Make a suspension with this bacterial colony in a steril tube of physiological water (5mL). In the water, rub inoculation loop on the tube side to disperse all the bacteria. WARNING! The sample is realized in asepsis around the Bunsen burner. 1.3. Repeat 4 times, in order to introduce 5 bacterial colonies in the tube

6. 1.4Homogenize your tube by manual rotations and write « I » on this tube. I

7. 2.Inoculate Mueller Hinton medium for antibiogram

8. 2.1. In the inoculum tube “I”, insert a sterile swab. WARNING! The sample is realized in asepsis around the Bunsen burner. Wet the swab and then wring the excess of liquid on the wall of the tube.

9. 2.2. Inoculate Mueller Hinton agar by rubbing the swab on its surface. Achieve tight streaks. 2.3. Reproducing the operation twice by turning 60° the box each time in order to ensure an good distribution of the inoculum on the surface of the medium : 60° = Streaks in 3 passes

10. 2.Filing the antibiotics disks

11. 3.1. Position the Mueller Hinton agar previously sown on the deposit plan provided 3.2. With a sterile forceps, place the antibiotics disks on the surface of the agar When filing disks, apply slight pressure ton ensure the maintenance of the disk, but not too much to avoid the risk of perforating the agar

12. 4.Medium incubation Incubate the box, in the inverted position, 16 h at 37°C minimum

13. 5.Analyse your results

14. 5.1. After incubation, measure the diameter of the growth inhibition zones around the antibiotics disks

15. 5.2. Use the reading table, to conclude on the sensitivity or on the resistance of the strain to different antibiotics tested Antibiotic testedResistance if the diameter of inhibition is less than : Sensitive if the diameter of inhibition is greater than : Ampicillin14 mm19 mm SXT10 mm16 mm Erythromycin17 mm22 mm Gentamicin14 mm16 mm

16. 6.Conclusion Suggest an antibiotic suitable for the treatment of nosocomial infection caused by your bacteria