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134 protocols using mic test strip

1

Evaluating Antibiotic Combination Efficacy against CR K. pneumoniae

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The in vitro effect of two combinations (ie, meropenem/ciprofloxacin and meropenem/colistin) was evaluated for CR K. pneumoniae isolates by “Perpendicular MIC test strip” method.16 (link)
MIC values of meropenem, ciprofloxacin, and colistin were determined by MIC test strip (Liofilchem, Italy) separately and in combinations and the fractional inhibitory concentration index (Σ FIC) was calculated for each combination using the formula: Σ FIC = FIC [A]+ FIC [B].
FIC [A]= MIC of drug A in combination with drug B/ MIC of drug A alone.
FIC [B]= MIC of drug B in combination with drug A/ MIC of drug B alone.
The Σ FIC values determine the effect of the tested combination as follows: ≤0.5, synergy; >0.5 to ≤1.0, additively; >1.0 to <4.0, indifference; and ≥4, antagonism.17
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2

Antibiotic Susceptibility of Lactobacillus Strains

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Bacteriostatic susceptibility was performed using Liofilchem MIC test strip (Liofilchem S.R.I., Italy) for chloramphenicol, erythromycin, sulfamethoxazole, tetracycline and trimethoprim according to the manufacturers’ instructions. The strips were placed on a Luria Berthani (LB) Agar inoculated with 0.5 ml LB pure colonies’ suspension of LpNC8 and LpNIZO2877, separately. Three independent replicates were performed for each antibiotic. Subsequently, the agar plates were incubated at 37 °C and observed for Minimum Inhibitory Concentration (MIC) reading after 24 h. MIC value was determined at the intersection of the strip and the growth inhibition ellipse, as reported by Kowalska-Krochmal et al.71 (link).
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3

Antimicrobial Susceptibility Testing of Salmonella Choleraesuis

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S. Choleraesuis isolates were tested for susceptibility to antimicrobial agents using the agar disk diffusion method (Kirby Bauer test) following the guidelines and criteria of the Clinical and Laboratory Standards Institute (CLSI) [26 ]. The antibiotics tested (Oxoid, England) were ampicillin (10 µg), amoxicillin–calvulanic acid (20/10 µg), aztreonam (30 µg), cefepime (30 µg), ceftazidime (30 µg), cefoxitin (30 µg), cefotaxime (30 µg), cefpodoxime (10 µg), cephalothin (30 µg), cefuroxime (30 µg), ceftriaxone (30 µg), ceftiofur (30 µg), ciprofloxacin (5 µg), chloramphenicol (30 µg), nalidixic acid (30 µg), norfloxacin (10 µg), streptomycin (10 µg), tetracycline (30 µg), and trimethoprim/sulfamethoxazole (25 µg). Furthermore, cefotaxime-resistant S. Choleraesuis isolates were screened for ESBL producers by the combination disk method (CLSI, 2008) [12 (link)]. The MICs for ciprofloxacin (32–0.002 µg/mL), cefotaxime (256-0.016 µg/mL), ceftazidime (256-0.016 µg/mL), ceftriaxone (256-0.016 µg/mL) and cefpodoxime (256-0.016 µg/mL) were determined using the Liofilchem MIC test strip (Liofilchem, Roseto degli Abruzzi, Italy), according to the manufacturer’s instructions. Escherichia coli ATCC 25922 was used as the quality control strain for antimicrobial susceptibility testing.
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4

Antimicrobial Susceptibility Testing Protocol

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Susceptibilities to ampicillin, azithromycin, clindamycin, cefoxitin, ciprofloxacin, daptomycin, erythromycin, gentamicin, linezolid, mupirocin, oxacillin, penicillin, rifampicin, quinupristin-dalfopristin, trimethoprim-sulfamethoxazole, teicoplanin, and vancomycin were tested using microdilution panels (MicroScan, Beckman Coulter Inc., Brea, CA, USA). Ceftaroline and telavancin MIC values were determined using the MIC test strip (Liofilchem, Roseto degli Abruzzi, Italy) and the Oxoid M.I.C.Evaluator™ (M.I.C.E™; Thermo Fisher Scientific, Basingstoke, England), respectively. The plates were incubated at 35 °C and read after 48 h. Population analysis was performed according to a reported method (Saito et al. 2014 (link)). Briefly, about 107–108 colony-forming units (CFU) of the overnight culture of each strain were inoculated on TSA plates containing various concentrations of vancomycin. The numbers of colonies formed after 48 h incubation at 35 °C were counted and plotted in a semilogarithmic graph.
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5

Vancomycin MIC Evaluation: A Standardized Protocol

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In this investigation, vancomycin MIC strips (MIC Test Strip, Liofilchem, Roseto degli Abruzzi Italy) were used. Serial microdilutions of native manufactured vancomycin (500 mg/vial, Dana Pharmacological Co, Tabriz, Iran) were performed, while Mueller-Hinton broth (Merck, Darmstadt, Germany), defibrinated lysed sheep blood (Baharafshan Co., Teheran, Iran), 96-well microplate (Technogen Spa, Pontenure Piacenza, Italy) and injectable distilled water were used.
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6

Antibiotic Resistance Testing of C. difficile

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Antibiotic resistance testing was performed with C. difficile suspensions in 0.9% NaCl at an optical density OD600 of 0.1 on Mueller Hinton agar enriched with 5% sheep blood at an anaerobic atmosphere with E-tests “MIC Test Strip” (Liofilchem®, Roseto degli Abruzzi, Italy) with metronidazole, vancomycin, erythromycin, moxifloxacin and rifampicin as described by the manufacturer. Applied resistance definitions were as follows: MIC (minimum inhibitory concentration) >2.0 µg/mL for vancomycin according to EUCAST (European Committee on Antimicrobial Susceptibility Testing) version 10.0, MIC > 2 µg/mL for metronidazole according to EUCAST version 10.0, MIC > 8 µg/mL for moxifloxacin according to CLSI (Clinical and Laboratory Standards Institute, M100S, 26th ed. 2016), MIC > 4 µg/mL for erythromycin according to DIN (“Deutsches Institut für Normierung”) version DIN 58940-1:2002-10 and MIC > 16 µg/mL for rifampicin according to [21 (link)].
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7

Vancomycin-Resistant E. faecium Characterization

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The 15 vancomycin-resistant E. faecium strains examined in this study were taken from a collection of non-consecutive clinical isolates identified at Geneva university hospitals (HUG) and by the Swiss National Reference Centre for Emerging Antibiotic Resistance (NARA). Table 1 depicts the characteristics of these clinical strains. Vancomycin MICs were determined by the gradient strip method (Etest, bioMérieux Marcy l'Etoile, France) according to the manufacturer's instructions. Importantly, the gradient strip method (Etest) or MIC Test Strip (Liofilchem, Italy) underestimates the vancomycin MIC for E. faecalis and E. faecium strains carrying the vanA/vanB genes but exhibiting low-level vancomycin resistance (Eucast, 2018 ). The identification of the strains was systematically confirmed by matrix-assisted desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS; compass, Bruker Daltonics, Bremen, Germany) according to the manufacturer's instructions.
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8

Comprehensive Antimicrobial Resistance Profiling

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EUCAST standardized DD testing and confirmation of species identification was performed on all isolates.5 ,15 (link) Broth microdilution (BMD) was performed on a representative proportion (every fourth E. coli, every third S. aureus and all isolates of the other species) of the NE isolates.4 ,14
E. coli and K. pneumoniae isolates resistant to cefotaxime and/or ceftazidime were further tested for ESBL [discs from Mast Diagnostics (Merseyside, UK)] and AmpC [MIC Test Strip (Liofilchem, Roseto degli Abruzzi, Italy)].16 Isolates resistant to either or both of the cephalosporins but phenotypically negative for ESBL and AmpC and isolates with a meropenem zone <28 mm with the reference method (the carbapenemase screening breakpoint of EUCAST)16 were sent to the Public Health Agency of Sweden for further characterization of resistance mechanisms using WGS.
All S. aureus isolates were screened for methicillin resistance with EUCAST standard DD using cefoxitin.14 ,16 The presence of mecA or mecC in screen-positive isolates was confirmed using standard molecular methods.
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9

Antibiotic Susceptibility Testing of E. coli Isolates

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Bacterial isolates from positive blood cultures were initially grown on blood- and MacConkey agar, while direct antimicrobial susceptibility testing (AST) from blood culture bottles was performed on MH-agar using the disk diffusion method and interpreted according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints [19 ]. In cases where no growth was observed on MH-agar, 5–7 colonies from blood agar were collected for alternative AST on blood agar, with E. coli CCUG 17620 as a susceptible control strain. All E. coli Isolates except from the first and the sixth BSI episodes were originally sent to the Norwegian National Advisory Unit on Detection of Antimicrobial Resistance (K-res) for confirmation of ESBL. Repeated AST of TMP-SMX for all isolates was subsequently performed with MIC Test Strip (Liofilchem) on MH-agar, blood agar and M9 agar medium added thymine (0.5, 2, 5 and 20 μg/mL).
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10

Determination of Antibiotic MIC Values

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MIC values were determined using MIC strips (MIC Test Strip, Liofilchem, Roseto degli Abruzzi, Italy) in LB plates at pH 4.6. Because no cefotiam MIC strips are commercially available, MIC values for this antibiotic were determined in 96-well microplates by serial dilutions of the antibiotic in liquid LB pH 4.6 medium and overnight incubation at 37°C. The starting inoculum was 6 × 105 cfu per well.
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