scholarly journals In VitroActivity of Fosfomycin against a Collection of Clinical Pseudomonas aeruginosa Isolates from 16 Spanish Hospitals: Establishing the Validity of Standard Broth Microdilution as Susceptibility Testing Method

2013 ◽  
Vol 57 (11) ◽  
pp. 5701-5703 ◽  
Author(s):  
María Díez-Aguilar ◽  
María-Isabel Morosini ◽  
Rosa del Campo ◽  
María García-Castillo ◽  
Javier Zamora ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Susceptibility Testing ◽  
Disk Diffusion ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Broth Microdilution ◽  
Content Type ◽  
Testing Method ◽  
Broth Microdilution Method ◽  
Agar Dilution

ABSTRACTThe broth microdilution method for fosfomycin andPseudomonas aeruginosawas assessed and compared with the approved agar dilution method in 206 genetically unrelatedP. aeruginosaclinical isolates. Essential agreement between the two methods was 84%, and categorical agreement was 89.3%. Additionally, Etest and disk diffusion assays were performed. Results validate broth microdilution as a reliable susceptibility testing method for fosfomycin againstP. aeruginosa. Conversely, unacceptable concordance was established between Etest and disk diffusion results with agar dilution results.

scholarly journals Comparison of Broth Microdilution Method UsingHaemophilus Test Medium and Agar Dilution Method for Susceptibility Testing of Eikenella corrodens

1998 ◽  
Vol 36 (8) ◽  
pp. 2386-2388 ◽  
Author(s):  
Luis Alcalá ◽  
Fernando García-Garrote ◽  
Emilia Cercenado ◽  
Teresa Peláez ◽  
Gema Ramos ◽  
...  
Keyword(s):  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Broth Microdilution ◽  
Test Medium ◽  
Eikenella Corrodens ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
Agar Dilution

Susceptibility testing of Eikenella corrodens is usually performed by a Mueller-Hinton sheep blood agar dilution (AD) method. However, this method is impractical for testing only a few strains. We compared AD with the broth microdilution method usingHaemophilus test medium (HTM) in order to determine the susceptibility of 36 clinical isolates of E. corrodens to eight antimicrobial agents. MICs obtained by the HTM method yielded 95.5 and 84% agreement (within 2 and 1 log2 dilutions, respectively) with those obtained by AD. The HTM method with incubation in CO2 for 48 h was highly reproducible and constitutes an easy alternative for antimicrobial susceptibility testing of E. corrodens.

scholarly journals Variation in erythromycin and clindamycin susceptibilities of Streptococcus pneumoniae by four test methods.

1997 ◽  
Vol 41 (1) ◽  
pp. 129-134 ◽  
Author(s):  
E L Fasola ◽  
S Bajaksouzian ◽  
P C Appelbaum ◽  
M R Jacobs
Keyword(s):  
Streptococcus Pneumoniae ◽  
Susceptibility Testing ◽  
Ambient Air ◽  
Test Methods ◽  
Disk Diffusion ◽  
Broth Microdilution ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
Agar Dilution ◽  
Resistant Strains

Susceptibilities of 124 strains of Streptococcus pneumoniae to erythromycin and clindamycin were determined by the National Committee for the Clinical Laboratory Standards (NCCLS) broth microdilution method, with incubation for 20 to 24 h in ambient air and with modifications of this method by incubation for up to 48 h in air and CO2. Strains were also tested by agar dilution, E-test, and disk diffusion; good correlation was obtained with these methods, with clear separation into bimodal populations of susceptible and resistant stains. The broth microdilution method, however, using incubation in air for 24 h (NCCLS method), misclassified 4 of 92 erythromycin-resistant strains (1 as susceptible and 3 as intermediate) and 25 of 58 clindamycin-resistant strains (all as susceptible). With the exception of one strain with clindamycin, susceptible and resistant strains were correctly classified by the microdilution method with incubation in CO2 for 24 h or in ambient air for 48 h. Disk diffusion, agar dilution, and E-test methods with incubation in 5% CO2 are therefore reliable methods for susceptibility testing of pneumococci against these agents. However, the NCCLS microdilution method, which specifies incubation for 20 to 24 h in ambient air, produced significant very major errors (43%) clindamycin. Modification of the microdilution method by incubation in 5% CO2 or by extension of incubation time in ambient air to 48 h corrected these errors. Disk diffusion, however, was shown to be a simple, convenient, and reliable method for susceptibility testing of pneumococci to erythromycin and clindamycin and is suggested as the method of choice for these agents.

scholarly journals Inaccuracy of the Disk Diffusion Method Compared with the Agar Dilution Method for Susceptibility Testing of Campylobacter spp.

2011 ◽  
Vol 50 (1) ◽  
pp. 52-56 ◽  
Author(s):  
Mirva Lehtopolku ◽  
Pirkko Kotilainen ◽  
Pauli Puukka ◽  
Ulla-Maija Nakari ◽  
Anja Siitonen ◽  
...  
Keyword(s):  
Susceptibility Testing ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Disk Diffusion Method ◽  
Agar Dilution ◽  
Campylobacter Spp

scholarly journals Broth Microdilution and Gradient Diffusion Strips vs. Reference Agar Dilution Method: First Evaluation for Clostridiales Species Antimicrobial Susceptibility Testing

Antibiotics ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 975
Author(s):  
Florian Baquer ◽  
Asma Ali Sawan ◽  
Michel Auzou ◽  
Antoine Grillon ◽  
Benoît Jaulhac ◽  
...  
Keyword(s):  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Reference Method ◽  
Antimicrobial Susceptibility Testing ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Broth Microdilution ◽  
Multicenter Studies ◽  
Gradient Diffusion ◽  
Agar Dilution

Antimicrobial susceptibility testing of anaerobes is challenging. Because MIC determination is recommended by both CLSI and EUCAST, commercial broth microdilution and diffusion strip tests have been developed. The reliability of broth microdilution methods has not been assessed yet using the agar dilution reference method. In this work, we evaluated two broth microdilution kits (MICRONAUT-S Anaerobes® MIC and Sensititre Anaerobe MIC®) and one gradient diffusion strip method (Liofilchem®) for antimicrobial susceptibility testing of 47 Clostridiales isolates (Clostridium, Clostridioides and Hungatella species) using the agar dilution method as a reference. The evaluation focused on comparing six antimicrobial molecules available in both microdilution kits. Analytical performances were evaluated according to the Food and Drug Administration (FDA) recommendations. Essential agreements (EA) and categorical agreements (CA) varied greatly according to the molecule and the evaluated method. Vancomycin had values of essential and categorical agreements above 90% for the three methods. The CA fulfilled the FDA criteria for three major molecules in the treatment of Gram-positive anaerobic infections (metronidazole, piperacillin/tazobactam and vancomycin). The highest rate of error was observed for clindamycin. Multicenter studies are needed to further validate these results.

scholarly journals Variability in Azithromycin Susceptibility Results for Neisseria gonorrhoeae Obtained Using Gradient MIC Strip and Agar Dilution Techniques

2019 ◽  
Vol 57 (12) ◽  
Author(s):  
Gary N. McAuliffe ◽  
Marian Smith ◽  
Gavin Cooper ◽  
Rose F. Forster ◽  
Sally A. Roberts
Keyword(s):  
Neisseria Gonorrhoeae ◽  
Susceptibility Testing ◽  
Geometric Mean ◽  
Test Strip ◽  
Clinical Isolates ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Content Type ◽  
Test Strips ◽  
Agar Dilution

ABSTRACT Azithromycin is a component of empirical treatment regimens for Neisseria gonorrhoeae infections, but antimicrobial susceptibility testing for this agent is technically challenging. We compared the intertest variability, MIC values, and CLSI/EUCAST categorization of clinical and reference isolates of N. gonorrhoeae treated with azithromycin by testing 107 clinical isolates and nine reference isolates by agar dilution and in duplicates using MIC test strips (Liofilchem, Italy) and Etests (bioMérieux, France). Replicate isolate agreement within 1 log2 between duplicate tests was 87% for MIC test strips and 100% for Etests (P < 0.001). Essential agreement with the agar dilution method was higher for Etests (91%) than for MIC test strips (44%, P < 0.001). The geometric mean MIC was highest for MIC test strips (0.8 mg/liter) and significantly higher than both Etest (0.47 mg/liter, P < 0.001) and agar dilution (0.26 mg/liter, P < 0.001) methods. Etest MICs were higher than those obtained with agar dilution (P < 0.001). Agar dilution, MIC test strip, and Etest methods categorized 96%, 85%, and 95% (P = 0.003) of clinical isolates, respectively, as susceptible/wild type according to CLSI/EUCAST criteria. Our results illustrate the difficulties underlying azithromycin susceptibility testing for N. gonorrhoeae and demonstrate that results can vary using different methods. This variability could influence antimicrobial resistance reporting between laboratories involved in N. gonorrhoeae surveillance programs.

scholarly journals Baseline Activity of Telavancin against Gram-Positive Clinical Isolates Responsible for Documented Infections in U.S. Hospitals (2011-2012) as Determined by the Revised Susceptibility Testing Method

2014 ◽  
Vol 59 (1) ◽  
pp. 702-706 ◽  
Author(s):  
Rodrigo E. Mendes ◽  
David J. Farrell ◽  
Helio S. Sader ◽  
Robert K. Flamm ◽  
Ronald N. Jones
Keyword(s):  
Staphylococcus Aureus ◽  
Susceptibility Testing ◽  
The United States ◽  
Broth Microdilution ◽  
Content Type ◽  
Testing Method ◽  
Microdilution Method ◽  
Baseline Activity ◽  
Broth Microdilution Method ◽  
Vancomycin Resistant

ABSTRACTTelavancin had MIC50and MIC90values of 0.03 and 0.06 μg/ml (100.0% susceptible), respectively, against methicillin-resistant and -susceptibleStaphylococcus aureus. Telavancin was active against vancomycin-susceptibleEnterococcus faecalis(MIC50/90, 0.12/0.12 μg/ml; 100% susceptible) andEnterococcus faecium(MIC50/90, 0.03/0.06 μg/ml), while higher MIC values were obtained against vancomycin-resistantE. faecium(MIC50/90, 1/2 μg/ml) andE. faecalis(MIC50/90, >2/>2 μg/ml). Streptococci showed telavancin modal MIC results of ≤0.015 μg/ml, except againstStreptococcus agalactiae(i.e., 0.03 μg/ml). This study reestablishes the telavancin spectrum of activity against isolates recovered from the United States (2011-2012) using the revised broth microdilution method.

scholarly journals Comparison of Agar Diffusion Methodologies for Antimicrobial Susceptibility Testing of Pseudomonas aeruginosa Isolates from Cystic Fibrosis Patients

2000 ◽  
Vol 38 (5) ◽  
pp. 1818-1822 ◽  
Author(s):  
Jane L. Burns ◽  
Lisa Saiman ◽  
Susan Whittier ◽  
Davise Larone ◽  
Jay Krzewinski ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Pseudomonas Aeruginosa ◽  
Susceptibility Testing ◽  
Correlation Coefficients ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Broth Microdilution ◽  
Disk Diffusion Method ◽  
Agar Diffusion ◽  
Broth Microdilution Method

Pseudomonas aeruginosa is the most common pathogen infecting the lungs of patients with cystic fibrosis (CF). Improved antimicrobial chemotherapy has significantly increased the life expectancy of these patients. However, accurate susceptibility testing of P. aeruginosa isolates from CF sputum may be difficult because the organisms are often mucoid and slow growing. This study of 597 CF isolates of P. aeruginosa examined the correlation of disk diffusion and Etest (AB BIODISK, Solna, Sweden) results with a reference broth microdilution method. The rates of interpretive errors for 12 commonly used antipseudomonal antimicrobials were determined. The disk diffusion method correlated well (zone diameter versus MIC) for all of the agents tested. However, for mucoid isolates, correlation coefficients (r values) for piperacillin, piperacillin-tazobactam, and meropenem were <0.80. The Etest correlation with reference broth microdilution results (MIC versus MIC) was acceptable for all of the agents tested, for both mucoid and nonmucoid isolates. Category interpretation errors were similar for the disk diffusion and Etest methods with 0.4 and 0.1%, respectively, very major errors (false susceptibility) and 1.1 and 2.2% major errors (false resistance). Overall, both agar diffusion methods appear to be broadly acceptable for routine clinical use in susceptibility testing of CF isolates of P. aeruginosa.

scholarly journals Revised Reference Broth Microdilution Method for Testing Telavancin: Effect on MIC Results and Correlation with Other Testing Methodologies

2014 ◽  
Vol 58 (9) ◽  
pp. 5547-5551 ◽  
Author(s):  
David J. Farrell ◽  
Rodrigo E. Mendes ◽  
Paul R. Rhomberg ◽  
Ronald N. Jones
Keyword(s):  
Susceptibility Testing ◽  
Previous Method ◽  
Broth Microdilution ◽  
Content Type ◽  
Testing Method ◽  
Microdilution Method ◽  
Drug Potency ◽  
Broth Microdilution Method ◽  
Plastic Surfaces

ABSTRACTThe reference broth microdilution (BMD) antimicrobial susceptibility testing method for telavancin was revised to include dimethyl sulfoxide (DMSO) as a solvent and diluent for frozen-form panel preparation, following the CLSI recommendations for water-insoluble agents. Polysorbate 80 (P-80) was also added to the test medium to minimize proven drug losses associated with binding to plastic surfaces. Four hundred sixty-two Gram-positive isolates, including a challenge set of organisms with reduced susceptibilities to comparator agents, were selected and tested using the revised method for telavancin, and the MIC results were compared with those tested by the previously established method and several Sensititre dry-form BMD panel formulations. The revised method provided MIC results 2- to 8-fold lower than the previous method when tested against staphylococci and enterococci, resulting in MIC50values of 0.03 to 0.06 μg/ml for staphylococci and 0.03 and 0.12 μg/ml forEnterococcus faeciumandEnterococcus faecalis, respectively. Less-significant MIC decreases (1 to 2 log2dilution steps) were observed when testing streptococci in broth supplemented with blood, which showed similar MIC50values for both methods. However,Streptococcus pneumoniaehad MIC50results of 0.008 and 0.03 μg/ml when tested by the revised and previous methods, respectively. Highest essential agreement rates (≥94.0%) were noted for one candidate dry-form panel formulation compared to the revised test. The revised BMD method provides lower MIC results for telavancin, especially when tested against staphylococci and enterococci. This is secondary to the use of DMSO for panel production and the presence of P-80, which ensure the proper telavancin testing concentration and result in a more accurate MIC determination. Moreover, earlier studies where the previous method was applied underestimated thein vitrodrug potency.

scholarly journals Performance of Four Fosfomycin Susceptibility Testing Methods against an International Collection of Clinical Pseudomonas aeruginosa Isolates

2020 ◽  
Vol 58 (10) ◽  
Author(s):  
Elizabeth C. Smith ◽  
Hunter V. Brigman ◽  
Jadyn C. Anderson ◽  
Christopher L. Emery ◽  
Tiffany E. Bias ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Susceptibility Testing ◽  
Wide Spectrum ◽  
Multidrug Resistant ◽  
Supporting Evidence ◽  
Broth Microdilution ◽  
Testing Methods ◽  
Content Type ◽  
E Coli ◽  
Agar Dilution

ABSTRACT Fosfomycin has been shown to have a wide spectrum of activity against multidrug-resistant Gram-negative bacteria; however, breakpoints have been established only for Escherichia coli or Enterobacterales per the Clinical and Laboratory Standards Institute (CLSI) and the European Committee on Antimicrobial Susceptibility Testing (EUCAST), respectively. A lack of additional organism breakpoints limits clinical use of this agent and has prompted extrapolation of these interpretive categories to other organisms like Pseudomonas aeruginosa without supporting evidence. Further complicating the utility of fosfomycin is the specified method for MIC determination, namely, agar dilution, which is not widely available and is both labor and time intensive. We therefore sought to determine the susceptibility of a large international collection of P. aeruginosa isolates (n = 198) to fosfomycin and to compare testing agreement rates across four methods: agar dilution, broth microdilution, disk diffusion, and Etest. Results were interpreted according to CLSI E. coli breakpoints, with 49.0 to 85.8% considered susceptible, dependent upon the testing method used. Epidemiological cutoff values were calculated and determined to be 256 μg/ml and 512 μg/ml for agar dilution and broth microdilution, respectively. Agreement rates were analyzed using both agar dilution and broth microdilution with a resulting high essential agreement rate of 91.3% between the two susceptibility testing methods. These results indicate that broth microdilution may be a reliable method for fosfomycin susceptibility testing against P. aeruginosa and stress the need for P. aeruginosa-specific breakpoints.

scholarly journals Comparative Analysis of Gradient Diffusion and Disk Diffusion with Agar Dilution for Susceptibility Testing of Elizabethkingia anophelis

Antibiotics ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 450
Author(s):  
Chien-Tung Chiu ◽  
Chung-Hsu Lai ◽  
Yi-Han Huang ◽  
Chih-Hui Yang ◽  
Jiun-Nong Lin
Keyword(s):  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Disk Diffusion Method ◽  
Gradient Diffusion ◽  
Minimum Requirements ◽  
Agar Dilution

Elizabethkingia anophelis has recently emerged as a cause of life-threatening infections. This study compared the results of antimicrobial susceptibility testing (AST) conducted for E. anophelis through different methods. E. anophelis isolates collected between January 2005 and June 2019 were examined for their susceptibility to 14 antimicrobial agents by using disk diffusion, gradient diffusion (Etest; (bioMérieux S.A., Marcy l’Etoile, France), and agar dilution methods. The agar dilution method was the reference assay. According to the agar dilution method, the isolates exhibited the highest susceptibility to minocycline (100%), doxycycline (97.6%), rifampin (95.2%), and levofloxacin (78.6%). A very major error rate of >1.5% was observed for nine antibiotics tested using the disk diffusion method. The overall categorical agreement rate between the disk diffusion and agar dilution methods was 74.8%, and ceftazidime, minocycline, levofloxacin, and rifampin met the minimum requirements for discrepancy and agreement rates. The Etest method tended to produce lower log2 minimum inhibitory concentrations for the antibiotics, except for trimethoprim–sulfamethoxazole and rifampin; the method resulted in very major errors for nine antibiotics. The overall essential and categorical agreement rates between the Etest and agar dilution methods were 67.3% and 76.1%, respectively. The Etest method demonstrated acceptable discrepancy and agreement rates for ceftazidime, minocycline, doxycycline, levofloxacin, and rifampin. AST results obtained through the disk diffusion and Etest methods for multiple antibiotics differed significantly from those obtained using the agar dilution method. These two assays should not be a routine alternative for AST for E. anophelis.

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