Antimicrobial susceptibility assay was performed on all staphylococci according to the Clinical and Laboratory Standards Institute guidelines [44 ]. Sixteen antimicrobial agents were utilized for disc diffusion assays on Mueller–Hinton agar (MHA, Difco Laboratories): ampicillin (AMP, 10 μg), cefoxitin (FOX, 30 μg), penicillin (PEN, 10 μg), chloramphenicol (CHL, 30 μg), ciprofloxacin (CIP, 5 μg), enrofloxacin (ENR, 5 μg), levofloxacin (LEV, 5 μg), clindamycin (CLI, 2 μg), erythromycin (ERY, 15 μg), gentamicin (GEN, 10 μg), mupirocin (MUP, 200 μg), rifampicin (RIF, 5 μg), sulfamethoxazole-trimethoprim (SXT, 23.75/1.25 μg), quinupristin-dalfopristin (SYN, 15 μg), and tetracycline (TET, 30 μg). mupirocin was purchased from Oxoid (Hampshire, UK), and the rest of the antimicrobial agents were purchased from BD BBLTM (Becton Dickinson, Franklin Lakes, NJ). The minimum inhibitory concentrations (MICs) of oxacillin (OXA), TET, CIP, and ENR were determined for all the study strains by using the standard two-fold broth microdilution [44 ]. The MICs of the study strains to vancomycin (VAN) and linezolid (LZD) were determined by standard Etest (AB Biodisk, Dalvagen, Sweden). S. aureus MW2 and S. aureus ATCC 29213 strains were used as reference strains for the antimicrobial susceptibility tests. All antimicrobial susceptibility tests were repeated three times.
Vancomycin
Manufactured by bioMérieux
Sourced in France, Sweden
Vancomycin is a diagnostic instrument developed by bioMérieux for the detection and quantification of vancomycin in biological samples. The instrument utilizes advanced analytical techniques to provide accurate and reliable results for healthcare professionals.
Automatically generated - may contain errors
Lab products found in correlation
Teicoplanin, by bioMérieux (6 mentions)
Daptomycin, by bioMérieux (6 mentions)
Etest, by bioMérieux (5 mentions)
Tetracycline, by bioMérieux (5 mentions)
Ciprofloxacin, by BD (4 mentions)
Cefoxitin, by bioMérieux (4 mentions)
Metronidazole, by bioMérieux (4 mentions)
Erythromycin, by bioMérieux (4 mentions)
Mupirocin, by BD (3 mentions)
Trimethoprim sulfamethoxazole, by BD (3 mentions)
Penicillin, by BD (3 mentions)
Erythromycin, by BD (3 mentions)
Gentamicin, by BD (3 mentions)
Tetracycline, by BD (3 mentions)
Clindamycin, by BD (3 mentions)
Clindamycin, by bioMérieux (3 mentions)
Linezolid, by bioMérieux (3 mentions)
Amikacin, by bioMérieux (3 mentions)
Tigecycline, by bioMérieux (3 mentions)
Streptomycin, by bioMérieux (3 mentions)
26 protocols using vancomycin
1
Antimicrobial Susceptibility Profiling of Staphylococci
2
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).
3
Vancomycin Susceptibility of MRSA
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All S. aureus isolates were identified using standard methods. Methicillin resistance was confirmed by polymerase chain reaction (PCR) detection of the mecA gene. Vancomycin MICs of methicillin-resistant S. aureus (MRSA) isolates were determined by the Vancomycin Etest (AB Biodisk, Piscataway, NJ, USA) on Mueller-Hinton agar according to the manufacturer's instructions.
4
Antimicrobial Susceptibility Testing of Bacterial Isolates
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Antimicrobial susceptibility testing was performed using the disk diffusion method on Muller-Hinton agar (Difco, Detroit, MI, USA) according to the recommendation to the guidelines of the European Committee on Antimicrobial Susceptibility Testing (EUCAST, 2017) (http://www.eucast.org/ , accesed on 9 January 2017) [30 ]. A total of 17 antibiotics (Oxoid, Basingstoke, UK) were tested: oxacillin (OXA, 1 μg), cefoxitin (FOX, 30 μg), penicillin (P, 10 UI), gentamicin (CN, 10 μg), ciprofloxacin (CIP, 5 μg), erythromycin (E, 15 μg), clindamycin (DA, 2 μg), chloramphenicol (C, 30 μg), rifampin (RD, 5 μg), tetracycline (TE, 30 μg), vancomycin (VA, 30 μg), Teicoplanin (TEC, 30 μg), trimethoprim-sulfamethoxazole (SXT, 1.25/23.75 μg), quinupristin/dalfopristin (QDA, 15 μg), fosfomycin (FOS, 50 µg), linezolid (LNZ, 30 µg) and fusidic acid (FD, 10 µg).
The minimum inhibitory concentrations (MICs) for oxacillin and cefoxitin were determined by E-test (AB-biodisk, Dalvogen, Sweden) on Muller-Hinton agar (Difco, Detroit, MI, USA) and interpreted as recommended by EUCAST [30 ].
Isolates that presented resistance to three or more classes of antibiotics, other than beta-lactams, were classified as having a multidrug resistance profile [31 (link)].
The minimum inhibitory concentrations (MICs) for oxacillin and cefoxitin were determined by E-test (AB-biodisk, Dalvogen, Sweden) on Muller-Hinton agar (Difco, Detroit, MI, USA) and interpreted as recommended by EUCAST [30 ].
Isolates that presented resistance to three or more classes of antibiotics, other than beta-lactams, were classified as having a multidrug resistance profile [31 (link)].
5
Isolation and Identification of Enterococcus spp.
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Feces (a total of 25 g) were diluted 1:10 in Brain Heart Infusion (BHI, Difco Laboratories, Detroit, MI, USA) broth supplemented with sterile NaCl 6.5% and incubated at 37 ± 1°C for 18 to 24 h. Subsequently, 20 µL of the incubated broth was subcultured onto m-Enterococcus Agar (Difco Laboratories) for the selective isolation of Enterococcus spp. Simultaneously, for the selective isolation of vancomycin-resistant enterococci (VRE), 10 µL of the initial broth was subcultured on BHI broth supplemented with 2 mg/L vancomycin (Sigma-Aldrich, Saint Louis, MO, USA). After incubation (37 ± 1°C for 18 to 24 h), 20 µL was subcultured onto m-Enterococcus plates with vancomycin (6 mg/mL) and incubated at 37 ± 1°C for 18 to 24 h. Ten colonies per plate were selected based on colony morphology and subcultured onto Blood Agar (Columbia agar + 5% sheep blood, bioMérieux, Marcy-l’Etoile, France) for further identification of Efs and Efm by the simultaneous real-time PCR amplification of the mltF gene of Efs and the ddlA gene present in Efm (RTi-PCR1), as described elsewhere (16 (link)).
6
Antibiotic Susceptibility Testing of Staphylococcus aureus
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Antibacterial susceptibility testing was performed by the disc diffusion method 21 with the following antimicrobial disks (Oxoid, Basingstoke, UK): benzyl penicillin (2 U), cefoxitin (30 μg), kanamycin (30 μg), mupirocin (200 μg and 5 μg), gentamicin (10 μg), erythromycin (15 μg), clindamycin (2 μg), chloramphenicol (30 μg), tetracycline (10 μg), trimethoprim (2.5 μg), fusidic acid (10 μg), rifampicin (5 μg), ciprofloxacin (5 μg), teicoplanin (30 μg), vancomycin (30 μg) and linezolid (30 μg). Discs containing cadmium acetate (50 μg), propamidine isethionate (100 μg) and mercuric chloride (109 μg) were prepared in the laboratory. Minimum inhibitory concentration (MIC) for cefoxitin, vancomycin and teicoplanin were determined with E-test strips (AB BioMérieux, Marcy l'Etoile, France) according to the manufacturer's instructions. Staphylococcus aureus strain ATCC25923 was used as a quality control strain for susceptibility testing.
7
Antimicrobial Susceptibility Testing for C. difficile
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Bacterial suspensions and MIC gradient stripes containing erythromycin, vancomycin, metronidazole, tetracycline and clindamycin (bioMérieux, Marcy l'Etoile, France; Oxoid; Himedia) were plated onto BD™ Brucella Blood Agar with Hemin and Vitamin K1 (Becton Dickinson) following the manufacturer's instructions. According to the recommendations of the Clinical and Laboratory Institute (CLSI), the following breakpoints for resistance were used: erythromycin and clindamycin 8 mg/L, tetracycline 16 mg/L, metronidazole 32 mg/L (CLSI, 2021 ; Spigaglia et al., 2011 (link)). CLSI‐approved clinical breakpoints for vancomycin were not available. The reference strain C. difficile ATCC®700057 was used as quality control strain.
8
Antibiotic Susceptibility Profiling of Staphylococcus
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Susceptibility to penicillin G, gentamicin, rifampin, ciprofloxacin, clindamycin, erythromycin, chloramphenicol, tetracycline, linezolid, and vancomycin (National Institutes for Food and Drug Control, China) were tested by agar dilution method as described by Wiegand et al.23 In addition, the E‐test method was used to determine the MICs of all isolates for sulfamethoxazole/ trimethoprim (SXT) (bioMérieux, France). Mueller‐ Hinton agar (MHA) plates were inoculated by streaking the standardized inocula (0.5 McFarland, approximately 1.5 × 108 CFUs/mL) using a sterile swab. The SXT E‐test strips (bioMérieux, France) were placed on the plates, followed by incubation at 35°C for 16–20 h. The minimal inhibitory concentration (MIC) reading for both the E‐test and agar dilution methods was conducted independently by a senior experimenter, with the result confirmed by a second reader. The MIC results were interpreted according to the Clinical and Laboratory Standards Institute (CLSI) breakpoints for Staphylococcus spp.24 Staphylococcus aureus ATCC29213 was used as a quality control. MDR was defined as isolates resistant to ≥3 classes of non‐β‐lactam antimicrobials.
9
Antibiotic Resistance Profiling
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Resistance to a set of other agents was assessed with the use of the disc diffusion (DD) method and the E-test method. The former (by Oxoid) was applied for penicillin (P); amikacin (AK); gentamycin (CN); ciprofloxacin (CIP); levofloxacin (LEV); mupirocin (MUP); fusidic acid (FUS); tetracycline (TET), and the latter (by BioMerieux) for ceftaroline (CPT); vancomycin (VA); teicoplanin (TP); linezolid (LZD); daptomycin (DPC); tigecycline (TGC); and spectinomycin (SC), according to the EUCAST guidelines [28 ,51 ].
10
Antibiotic Susceptibility of Marseille-P3237
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The antibiotic susceptibility of strain Marseille-P3237 was assessed using the E-test method for the following molecules: benzylpenicillin, amoxicillin, cefotaxime, ceftriaxone, imipenem, amikacin, erythromycin, daptomycin, rifampicin, minocycline, teicoplanin, vancomycin, colistin and metronidazole (bioMérieux).
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