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6 protocols using fusidic acid

1

Antibiotic Susceptibility Testing of S. aureus

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S. aureus isolates obtained at C1 and C2 were suspended in sterile saline to 0.5 McFarland. Susceptibility testing was performed on Mueller-Hinton II agar 3.8% w/v (BD Diagnostic Systems, Sparks, MD, USA) using the standardized disk diffusion method in accordance with the European Committee on Antimicrobial Susceptibility Testing (www.eucast.org) guidelines for the following antibiotics: cefoxitin (30 µg), fusidic acid (10 µg), erythromycin (15 µg), clindamycin (2 µg), rifampicin (5 µg), gentamicin (10 µg), trimethoprim-sulfamethoxazole (25 µg), tetracycline (30 µg), and norfloxacin (10 µg). All disks were from Oxoid, Basingstoke, UK.
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2

Antimicrobial Susceptibility Testing of CA-MRSA

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All S. aureus isolates were identified by mass spectrometry (MALDI-TOF, Biotyper, Bruker Daltonic GmBH, Bremen, Germany). The susceptibility of CA-MRSA isolates was tested against 16 antimicrobial agents using the disk diffusion method according to the guidelines of the Clinical Laboratory Standard Institute (CLSI) (13 ). The antibiotics tested were penicillin, cefoxitin, vancomycin, gentamicin, tobramycin, kanamycin, erythromycin, clindamycin, tetracycline, ciprofloxacin, trimethoprimsulfamethoxazole, chloramphenicol, rifampin, linezolid, mupirocin and fusidic acid (BD, Maryland, USA). Minimal inhibitory concentration (MIC) determination of oxacillin was performed using the E-test (bioMerieux, Marcy I’Etoile, France).
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3

Antibiotic Susceptibility Profiling of MRSA

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The susceptibility patterns of the MRSA isolates were performed by the agar disk diffusion method according to the guidelines of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) (15 , 16 ). The antibiotics tested were penicillin, cefoxitin, gentamicin, tobramycin, kanamycin, erythromycin, clindamycin, tetracycline, ciprofloxacin, trimethoprim-sulfamethoxazole, chloramphenicol, rifampin, linezolid, mupirocin and fusidic acid (BD, Sparks, USA). Minimal inhibitory concentration (MIC) determination of oxacillin, cefoxitin and vancomycin was performed using the E-test (bioMerieux, Marcy-l’Etoile, France).
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4

Antimicrobial Susceptibility of S. aureus

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The antimicrobial susceptibility of S. aureus strains was tested by disk diffusion method using penicillin G, erythromycin, clindamycin, ciprofloxacin, tetracycline, gentamicin, sulfamethoxazole/trimethoprim, chloramphenicol and fusidic acid discs from Becton Dickinson (Franklin Lakes, NJ, USA) and was interpreted according to the Clinical and Laboratory Standards Institute (CLSI).14 MIC for vancomycin was determined by E-tests, in line with the manufacturer’s instructions (AB Biodisc, Solna, Sweden).
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5

Antibiotic Resistance Profiling of S. aureus

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The resistance of S. aureus isolates to antimicrobial agents was determined by disk diffusion and interpreted according to the CLSI document no. M02-A11.14 The following drugs were used for the test: penicillin, erythromycin, azithromycin, roxithromycin, clindamycin, lincomycin, ciprofloxacin, ofloxacin, levofloxacin, tetracycline, amikacin, tobramycin, netilmicin, gentamicin, fusidic acid, sulfamethoxazole/trimethoprim, chloramphenicol, vancomycin (all from Becton Dickinson, Franklin Lakes, NJ, USA) and mupirocin (Oxoid). Multidrug resistance was defined as resistance to antimicrobial agents from at least three various classes. For isolates identified as resistant to erythromycin but susceptible to clindamycin, D-test was performed to detect inducible clindamycin resistance. Minimal inhibitory concentration (MIC) for vancomycin was determined by E-tests, in line with the manufacturer’s instructions (AB Biodisk, Solna, Sweden).
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6

Antimicrobial Resistance Profiling of Staphylococci

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Resistance to methicillin was primary tested using disc diffusion method on Mueller Hinton agar (Oxoid, UK), with oxacillin and cefoxitin discs (Becton Dickinson, Germany) and confirmed by the detection of mecA gene by polymerase chain reaction (PCR) (Barski et al. 1996 (link)). The susceptibility to antimicrobial agents was determined by the disc diffusion method according to recommendations given by Hryniewicz et al. (2005 ) and the CLSI guidelines (2006 ). The antibiotics tested were the following: penicillin, erythromycin, clindamycin, ciprofloxacin, co-trimoxazole, tetracycline, gentamicin, vancomycin, teicoplanin, fusidic acid, rifampicin, linezolid, quinupristin-dalfopristin, chloramphenicol (Becton Dickinson, Germany), and mupirocin (Oxoid, UK). For all the isolates beta-lactamase production was checked by nitrocefin test (Becton Dickinson, Germany). For isolates identified as resistant to erythromycin, but susceptible to clindamycin, D-test was performed to detect inducible clindamycin resistance (Fiebelkorn et al. 2003 (link)). The susceptibility to vancomycin was confirmed by the minimal inhibitory concentration (MIC) using E-tests, as described by the manufacturer (AB Biodisc, Sweden).
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