scholarly journals Daptomycin Bactericidal Activity and Correlation between Disk and Broth Microdilution Method Results in Testing of Staphylococcus aureus Strains with Decreased Susceptibility to Vancomycin

2006 ◽  
Vol 50 (7) ◽  
pp. 2330-2336 ◽  
Author(s):  
Helio S. Sader ◽  
Thomas R. Fritsche ◽  
Ronald N. Jones
Keyword(s):  
Staphylococcus Aureus ◽  
Bactericidal Activity ◽  
Correlation Coefficients ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Broth Microdilution ◽  
Disk Diffusion Method ◽  
Initial Inoculum ◽  
Microdilution Method ◽  
Broth Microdilution Method

ABSTRACT A total of 207 Staphylococcus aureus strains, including 105 well-characterized strains with decreased susceptibility to vancomycin (17 vancomycin-intermediate S. aureus [VISA] and 88 heteroresistant VISA [hVISA] strains) and 102 wild-type methicillin-resistant S. aureus (MRSA-WT) strains were tested by reference/standardized broth microdilution and disk diffusion methods, as well as by Etest (AB BIODISK, Solna, Sweden), against daptomycin and vancomycin. The lowest concentration of antimicrobial agent that killed ≥99.9% of the initial inoculum was defined as the minimum bactericidal concentration (MBC) endpoint, and time-kill curves were performed in selected strains to further evaluate bactericidal activity. All MRSA-WT and hVISA strains were inhibited by ≤1 μg/ml of daptomycin, while the VISA strains showed slightly higher daptomycin MICs (range, 0.5 to 4 μg/ml). All daptomycin MBC results were at the MIC or twofold higher. In contrast, 14.7% of MRSA-WT, 69.3% of hVISA, and all VISA strains showed a vancomycin MBC/MIC ratio of ≥32 or an MBC of ≥16 μg/ml (tolerant). The correlation coefficients between broth microdilution and disk diffusion method results were low for daptomycin (0.07) and vancomycin (0.11). Eight (3.8%) strains (all hVISA or VISA) were “nonsusceptible” to daptomycin by broth microdilution methods but susceptible by the disk diffusion method. For vancomycin, 35 (16.9%) strains were nonsusceptible by broth microdilution methods but susceptible by disk diffusion methods. In conclusion, daptomycin was highly bactericidal against S. aureus strains, and its bactericidal activity was not affected by decreased susceptibility to vancomycin. In contrast, many (one in seven) contemporary MRSA-WT, the majority of hVISA, and all VISA strains showed vancomycin MBC/MIC ratios consistent with tolerance, a predictor of poor clinical response. Disk diffusion tests generally failed to detect strains categorized as nonsusceptible to daptomycin or vancomycin by the reference broth microdilution method or Etest, and reassessment of breakpoints should be immediately attempted for MIC methods suggested as the test of choice.

scholarly journals Categorizing susceptibility of clinical isolates of Candida auris to amphotericin B, caspofungin, and fluconazole utilizing the CLSI M44-A2 disk diffusion method

2021 ◽  
Author(s):  
Natalie S. Nunnally ◽  
Tajah Damm ◽  
Shawn R. Lockhart ◽  
Elizabeth L. Berkow
Keyword(s):  
Amphotericin B ◽  
Accurate Method ◽  
Clinical Isolates ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Broth Microdilution ◽  
Candida Auris ◽  
Disk Diffusion Method ◽  
Microdilution Method ◽  
Broth Microdilution Method

We evaluated the CLSI M44ed3E disk diffusion method in comparison with the CLSI M27ed4 broth microdilution method for caspofungin and fluconazole and the Etest method for amphotericin B to categorize susceptibility of 347 clinical isolates of Candida auris. Utilizing the zone diameter cutoffs established here we observed the overall categorial agreement between the two methods. For caspofungin, concordant results were observed for 98% of isolates with <1% very major and 1% major errors. For fluconazole, concordant results were observed for 91% of isolates with 1% very major and 8% major errors. For amphotericin B, concordant results were observed for 74% of isolates with <1% very major errors and 25% major errors. The disk diffusion approach provides an accurate method for determining the susceptibility of C. auris for caspofungin and fluconazole, and for identification of at least 75% of amphotericin B-susceptible isolates.

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 Evaluation of Disk Diffusion Method for Determining Posaconazole Susceptibility of Filamentous Fungi: Comparison with CLSI Broth Microdilution Method

2006 ◽  
Vol 50 (3) ◽  
pp. 1108-1111 ◽  
Author(s):  
E. López-Oviedo ◽  
A. I. Aller ◽  
C. Martín ◽  
C. Castro ◽  
M. Ramirez ◽  
...  
Keyword(s):  
Filamentous Fungi ◽  
Clinical Laboratory ◽  
Culture Media ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Broth Microdilution ◽  
Disk Diffusion Method ◽  
Microdilution Method ◽  
Broth Microdilution Method

ABSTRACT The disk diffusion method was evaluated for determining posaconazole susceptibility against 78 strains of molds using two culture media in comparison with the CLSI (Clinical Laboratory Standards Institute) broth microdilution method (M38-A). A significant correlation between disk diffusion and microdilution methods was observed with both culture media.

scholarly journals Evaluation of the Revised Ceftaroline Disk Diffusion Breakpoints When Testing a Challenge Collection of Methicillin-ResistantStaphylococcus aureusIsolates

2018 ◽  
Vol 56 (12) ◽  
Author(s):  
Helio S. Sader ◽  
Paul R. Rhomberg ◽  
Timothy B. Doyle ◽  
Robert K. Flamm ◽  
Rodrigo E. Mendes
Keyword(s):  
Error Rates ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Broth Microdilution ◽  
Wild Type ◽  
Disk Diffusion Method ◽  
Content Type ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
Mueller Hinton Agar

ABSTRACTWe assessed ceftaroline disk diffusion breakpoints forStaphylococcus aureuswhen applying revised Clinical and Laboratory Standards Institute (CLSI) ceftaroline MIC breakpoints. Disk-MIC correlation was evaluated by testing a challenge collection (n= 158) of methicillin-resistantS. aureus(MRSA) isolates composed of 106 randomly selected isolates plus 52 isolates with decreased susceptibility to ceftaroline (MIC, 1 to 16 μg/ml). Disk diffusion was performed with 30-μg disks and Mueller-Hinton agar from 2 manufacturers each. Revised CLSI susceptible (S)/susceptible dose-dependent (SDD)/resistant (R) MIC breakpoints of ≤1/2 to 4/≥8 μg/ml were applied. The disk breakpoints that provided the lowest error rates were CLSI S/R breakpoints of ≥25 mm/≤19 mm, with no very major (VM) or major (Ma) errors and with minor (Mi) error rates of 0.0% for ≥2 doubling dilutions above the I or SDD (≥I + 2), 22.1% for I or SDD plus or minus 1 doubling dilution (I ± 1), and 2.3% for ≤2 doubling dilutions below the I or SDD ≤I − 2 (overall Mi error rate, 16.5%). No mutation in the penicillin-binding protein 2a (PBP2a) was observed in 5 of 15 isolates with a ceftaroline MIC of 2 μg/ml; 3 of 11 isolates with a ceftaroline MIC of 1 μg/ml exhibited mutations in the penicillin-binding domain (PBD; 1 isolate) or in the non-PBD (2 isolates). All isolates except 1, with a ceftaroline MIC of ≥4 μg/ml, showed ≥1 mutation in the PBD and/or non-PBD. In summary, results from the disk diffusion method showed a good correlation with those from the reference broth microdilution method. Our results also showed that the ceftaroline MIC distribution of isolates with no mutations in the PBP2a goes up to 4 μg/ml, and reference broth microdilution and disk diffusion methods do not properly separate wild-type from non-wild-type isolates.

scholarly journals Investigation of susceptibility of Staphylococcus species to some antibacterial drugs by disk diffusion and broth microdilution

2012 ◽  
Vol 66 (3-4) ◽  
pp. 199-210
Author(s):  
Jelena Asanin ◽  
Ksenija Aksentijevic ◽  
Milenko Zutic ◽  
Vera Katic ◽  
Dejan Krnjaic ◽  
...  
Keyword(s):  
Disk Diffusion ◽  
Diffusion Method ◽  
Broth Microdilution ◽  
Becton Dickinson ◽  
Disk Diffusion Method ◽  
Antibacterial Drugs ◽  
Milk Samples ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
Resistance To Penicillin

The objective of this work was to identify isolated Staphylococcus species and to investigate their sensitivity to some antibacterial drugs. The material used for these investigations were Staphylococcus isolates originating from milk samples. A total of 25 strains of Staphylococcus isolates were examined, including 24 from milk samples from cows with mastitis, and one strain was isolated from a milk sample from a cow following treatment for mastitis. For primary identification, catalase and oxidase tests were used, as well as the free coagulase test. Following the preliminary tests, the isolated strains were identified using commercial systems ID32 STAPH (bioM?rieux, France) and the BBL Crystal Gram-Positive ID Kit (Becton Dickinson, USA) according to the enclosed instructions. The Staphylococcus isolates were examined for sensitivity to the following: oxacillin, penicillin, cefoxitin, gentamicin, erythromycin, chloramphenicol, tetracycline, ciprofloxacin, sulfametoxazol/trimetoprim, and vacomycin using the disk diffusion method and the broth microdilution method as recommended by the Clinical and Laboratory Strandards Institute - CLSI(2003), and the results were interpreted according to CLSI recommendations from 2008 and 2010. Antibiogram disks manufactured by Becton Dickinson (USA) were used, and the broth microdilution method was applied using pure antibiotic substances from different manufacturers: erythromycin, chloramphenicol, cefoxitin, gentamicin, oxacillin, tetracycline (Sigma Aldrich, USA), sulfametoxazol (Fluka, USA), penicillin (Calbiochem, Germany), vancomycin (Abbott laboratories, USA), ciprofloxacin and trimetoprim (Zdravlje A.D., Serbia). All 25 strains were catalase positive and oxidase negative. Of the 25 strains, 19 were coagulase positive and 6 were coagulase negative.With the implementation of the disk diffusion method on 19 strains of S. aureus, 17 were established to be resistant to penicillin (89.5%), and 2 strains to gentamicin (10.5%). The investigation of 3 strains of S. xylosus using the disk diffusion method showed that one strain was resistant to tetracycline (33.3%) and to oxacillin (33.3%), while another strain was found to be resistant to penicillin (33.3%). The third strain of S. xylosus was sensitive to all the examined antibiotics. Two strains of S. simulans and one strain of S. haemolyticus were not found to be resistant to any of the examined antibiotics using the disk diffusion method. The implementation of the broth microdilution method yielded in 13 strains of S. aureus resistance to penicillin (68.4%) with MIC values from 0.5 to 4 ?g/m, in 2 strains to gentamicin (10.5%) with MIC values of 32 ?g/ml, and intermediary sensitivity to chloramphenicol was established in 9 strains of S. aureus (47.4%) with MIC values of 16 ?g/ml and to vancomycin in 1 strain of S. aureus (5.3%) with MIC values of 4 ?g/ml.

scholarly journals Evaluation of Identification and Susceptibility for Candida Spp. Isolated Directly from Positive Blood Culture Bottles

2021 ◽  
Vol 2021 ◽  
pp. 1-5
Author(s):  
Ândrea Celestino de Souza ◽  
Luciano Z. Goldani ◽  
Eliane Würdig Roesch ◽  
Larissa Lutz ◽  
Patricia Orlandi Barth ◽  
...  
Keyword(s):  
Blood Culture ◽  
Standard Method ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Identification System ◽  
Broth Microdilution ◽  
Disk Diffusion Method ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
Standard Disk

Determination of the susceptibility profile of isolates of Candida from blood culture bottles is extremely important for correctly guiding patient pharmacotherapy. The aim of this study was to compare the results of analysis of Candida isolated directly from blood culture bottles by the VITEK MS MALDI-TOF identification system and the fluconazole disk diffusion assay with those of standard identification methods. Testing directly from the bottle allowed results 24 to 48 hours quicker than the standard method. There was a categorical agreement of 51.64% (47 of 91 samples) between the results of analysis directly from the bottle and analysis by the standard method. Regarding species identification, there was 96.15% agreement for Candida parapsilosis (25 of 26 samples). Categorical agreement between the rapid and standard disk diffusion methods was 95%, and the agreement between the rapid disk diffusion method and the broth microdilution method was 97%. Only minor errors in the rapid method were observed: 3 (5%) in the standard disk diffusion method and 2 (3%) in the broth microdilution method. Our study concluded that the rapid disk diffusion method for fluconazole is a fast, easy, reproducible, and consistent method. Its timely implementation for testing antifungal agents in the clinical microbiology laboratory can help reduce profile release times, thus helping to determine the most appropriate antifungal treatment.

scholarly journals Antifungal Activity of Micafungin in Serum

2009 ◽  
Vol 53 (10) ◽  
pp. 4559-4562 ◽  
Author(s):  
Jun Ishikawa ◽  
Tetsuo Maeda ◽  
Itaru Matsumura ◽  
Masato Yasumi ◽  
Hidetoshi Ujiie ◽  
...  
Keyword(s):  
Antifungal Activity ◽  
High Performance ◽  
Diffusion Method ◽  
Broth Microdilution ◽  
Serum Samples ◽  
Disk Diffusion Method ◽  
Serum Free ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
Applied Dose

ABSTRACT We have evaluated the antifungal activity of micafungin in serum by using the disk diffusion method with serum-free and serum-added micafungin standard curves. Serum samples from micafungin-treated patients have been shown to exhibit adequate antifungal activity, which was in proportion to both the applied dose and the actual concentration of micafungin measured by high-performance liquid chromatography. The antifungal activity of micafungin in serum was also confirmed with the broth microdilution method.

scholarly journals Comparison of antimicrobial susceptibilities of Corynebacterium species by broth microdilution and disk diffusion methods.

1996 ◽  
Vol 40 (4) ◽  
pp. 930-933 ◽  
Author(s):  
K Weiss ◽  
M Laverdière ◽  
R Rivest
Keyword(s):  
Clinical Laboratory ◽  
High Rate ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
National Committee ◽  
Broth Microdilution ◽  
Disk Diffusion Method ◽  
Corynebacterium Species ◽  
Broth Microdilution Method

Corynebacterium species are increasingly being implicated in foreign-body infections and in immunocompromised-host infections. However, there are no specific recommendations on the method or the criteria to use in order to determine the in vitro activities of the antibiotics commonly used to treat Corynebacterium infections. The first aim of our study was to compare the susceptibilities of various species of Corynebacterium to vancomycin, erythromycin, and penicillin by using a broth microdilution method and a disk diffusion method. Second, the activity of penicillin against our isolates was assessed by using the interpretative criteria recommended by the National Committee for Clinical Laboratory Standards for the determination of the susceptibility of streptococci and Listeria monocytogenes to penicillin. Overall, 100% of the isolates were susceptible to vancomycin, while considerable variations in the activities of erythromycin and penicillin were noted for the different species tested, including the non-Corynebacterium jeikeium species. A good correlation in the susceptibilities of vancomycin and erythromycin between the disk diffusion and the microdilution methods was observed. However, a 5% rate of major or very major errors was detected with the Listeria criteria, while a high rate of minor errors (18%) was noted when the streptococcus criteria were used. Our findings indicate considerable variations in the activities of erythromycin and penicillin against the various species of Corynebacterium. Because of the absence of definite recommendations, important discrepancies were observed between the methods and the interpretations of the penicillin activity.

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