Accuracy of gradient diffusion method for susceptibility testing of dalbavancin and comparators

2021 ◽  
pp. 1-5
Author(s):  
A. G. Leroy ◽  
V. Lavigne-Quilichini ◽  
P. Le Turnier ◽  
B. Loufti ◽  
E. Le Breton ◽  
...  
Keyword(s):  
Susceptibility Testing ◽  
Diffusion Method ◽  
Gradient Diffusion

scholarly journals Multicenter Evaluation of the Etest Gradient Diffusion Method for Ceftolozane-Tazobactam Susceptibility Testing of Enterobacteriaceae and Pseudomonas aeruginosa

2018 ◽  
Vol 56 (9) ◽  
Author(s):  
Adam L. Bailey ◽  
Tom Armstrong ◽  
Hari-Prakash Dwivedi ◽  
Gerald A. Denys ◽  
Janet Hindler ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Susceptibility Testing ◽  
Clinical Isolates ◽  
Diffusion Method ◽  
Beta Lactam ◽  
Content Type ◽  
Gradient Diffusion ◽  
Tract Infections ◽  
Abdominal Infections ◽  
Inhibitor Combination

ABSTRACT Ceftolozane-tazobactam (C/T) is a novel beta-lactam–beta-lactamase inhibitor combination antibiotic approved by the U.S. Food and Drug Administration in 2014 for the treatment of complicated intra-abdominal infections (in combination with metronidazole) and complicated urinary tract infections. In this study, we evaluated the performance of the C/T Etest, a gradient diffusion method. C/T Etest was compared to broth microdilution (BMD) for 51 Enterobacteriaceae challenge isolates and 39 Pseudomonas aeruginosa challenge isolates at three clinical sites. Essential agreement (EA) between the methods ranged from 47 to 49/51 (92.2 to 96.1%) for the Enterobacteriaceae, and categorical agreement (CA) ranged from 49 to 51/51 (96.1 to 100.0%). EA and CA for P. aeruginosa were 100% at all sites. The C/T Etest was also compared to BMD for susceptibility testing on 966 clinical isolates (793 Enterobacteriaceae, including 167 Klebsiella pneumoniae and 159 Escherichia coli isolates, in addition to 173 P. aeruginosa isolates) collected at four clinical sites. EA between Etest and BMD was 96.9% for Enterobacteriaceae isolates and 98.8% for P. aeruginosa isolates. Within the Enterobacteriaceae, isolates from each species examined had >96% CA. For the clinical isolates, no very major errors were identified but two major errors were found (one for K. pneumoniae and one for Providencia rettgeri). By BMD, 47.0% of Enterobacteriaceae and 46.2% of P. aeruginosa challenge strains were nonsusceptible to C/T by CLSI breakpoint criteria; 8.2% of clinical Enterobacteriaceae isolates and 12.1% of clinical P. aeruginosa isolates were nonsusceptible to C/T by CLSI breakpoint criteria. In conclusion, Etest is accurate and reproducible for C/T susceptibility testing of Enterobacteriaceae and P. aeruginosa.

scholarly journals Multicenter Evaluation of the New Etest Gradient Diffusion Method for Piperacillin-Tazobactam Susceptibility Testing of Enterobacterales, Pseudomonas aeruginosa, and Acinetobacter baumannii Complex

2019 ◽  
Vol 58 (2) ◽  
Author(s):  
Sergio García-Fernández ◽  
Yohann Bala ◽  
Tom Armstrong ◽  
María García-Castillo ◽  
Carey-Ann D. Burnham ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Acinetobacter Baumannii ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Hospital Setting ◽  
Antimicrobial Susceptibility Testing ◽  
International Standards ◽  
Diffusion Method ◽  
Content Type ◽  
Gradient Diffusion

ABSTRACT Piperacillin-tazobactam (P/T) is a β-lactam–β-lactamase inhibitor combination frequently used in the hospital setting. Etest is a gradient diffusion method that represents an alternative to broth microdilution (BMD) for performing antimicrobial susceptibility testing. We conducted a multicenter evaluation of the performance of the new P/T Etest compared to that of BMD following U.S. Food and Drug Administration (FDA) and International Standards Organization (ISO) standard ISO 20776-2 criteria using Clinical and Laboratory Standards Institute (CLSI)-FDA and European Committee on Antimicrobial Susceptibility Testing (EUCAST) interpretive breakpoints, respectively. A total of 977 isolates (775 Enterobacterales isolates, 119 Pseudomonas aeruginosa isolates, and 83 Acinetobacter baumannii complex isolates) were tested. Overall essential agreement (EA) was 96.4% and 96.6% for Enterobacterales when FDA and ISO 20776-2 criteria, respectively, were followed. EA was 98.3% for P. aeruginosa and 91.6% for the A. baumannii complex when both the FDA and ISO criteria were followed. Applying CLSI-FDA breakpoints, categorical agreement (CA) reached 93.0%, 93.3%, and 89.2% for the Enterobacterales, P. aeruginosa, and the A. baumannii complex, respectively. Two very major errors (VMEs; 1.1%) were found among the Enterobacterales (for 2 Klebsiella pneumoniae isolates). No additional major errors (MEs) or VMEs were found. Applying EUCAST breakpoints, CA was 94.8% and 95.8% for Enterobacterales and P. aeruginosa, respectively (no breakpoints are currently available for the A. baumannii complex). No VMEs were observed among the Enterobacterales, but 2 (0.4%) MEs were found. Among the P. aeruginosa isolates, 2 (6.9%) VMEs and 3 (3.3%) MEs were observed. These errors resulted when P/T Etest MICs were 1 doubling dilution apart from the BMD MICs. In conclusion, the new P/T Etest represents an accurate tool for performing antimicrobial susceptibility testing of Enterobacterales, P. aeruginosa, and A. baumannii complex isolates with limited category errors.

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.

scholarly journals Validation of a Gradient Diffusion Method (Etest®) for Antimicrobial Susceptibility Testing of Aerococcus urinae to Fluoroquinolones

2021 ◽  
Author(s):  
France Emilie Roy ◽  
Tammy Berteau ◽  
Julie Bestman-Smith ◽  
Simon Grandjean Lapierre ◽  
Simon Frédéric Dufresne ◽  
...  
Keyword(s):  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Reference Method ◽  
Error Rates ◽  
Antimicrobial Susceptibility Testing ◽  
Diffusion Method ◽  
Sheep Blood ◽  
Gradient Diffusion ◽  
Reference Methods ◽  
Aerococcus Urinae

Aerococcus urinae is a urinary pathogen with well-described resistance to fluoroquinolones. This study aimed to validate the gradient diffusion (GD) method (Etest®) on cation-adjusted Mueller-Hinton agar with 5% sheep blood for Aerococcus urinae antimicrobial susceptibility testing (AST) to ciprofloxacin and levofloxacin and compare it to the broth microdilution (BMD) method from CLSI M45-A3. Agar dilution (AD), as recommended by EUCAST, was used as an alternate reference method to arbitrate discrepancies or address technical issues. Aerococcus urinae isolates from urinary specimens were prospectively collected between June 2016 and December 2017 from six Quebec hospitals (Canada) and identifications were confirmed using Vitek MS® with IVD 3.0 database. Of the 207 isolates tested using BMD, 37 (17.9%) showed trailing and 19 (9.2%) showed insufficient growth and were tested using AD. Also, 38 isolates (18.4%) for ciprofloxacin and 13 isolates (6.3%) for levofloxacin showed a lack of essential or categorical agreement between Etest® and BMD and were also tested by AD. Using a combined reference method (BMD or AD), susceptibility rate of Aerococcus urinae was 82.6% and 81.6% for ciprofloxacin and levofloxacin, respectively. Categorial agreement between GD and the combined reference methods was 95.2% for ciprofloxacin and 97.1% for levofloxacin, with no very major error identified. Major and minor error rates were 0.6% and 4.3% for ciprofloxacin, and 1.2% and 1.9% for levofloxacin, respectively. Overall, AST using Etest® on sheep blood agar showed a good agreement with reference methods and can be considered by clinical laboratories wishing to perform AST on Aerococcus urinae isolates.

scholarly journals Antimicrobial susceptibility patterns of anaerobic bacteria in Victoria, Australia

2018 ◽  
Author(s):  
Jasmine C. Teng ◽  
L Linda Dreyer
Keyword(s):  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Anaerobic Bacteria ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Clinical Diagnostic Laboratory ◽  
Diagnostic Laboratory ◽  
Gradient Diffusion ◽  
Anaerobic Infections ◽  
Susceptibility Patterns

AbstractMortality associated with anaerobic infections approximates 20%. Resistance of anaerobic bacteria to commonly used antimicrobials has been increasingly reported. The aim of this study was to describe antimicrobial susceptibility patterns of anaerobic bacteria isolated from clinical samples using a gradient diffusion method, E test (bioMérieux), in Victoria, Australia. Metronidazole, meropenem and amoxycillin-clavulanate were found to be active against almost all isolates tested. Most Gram positive anaerobic cocci (GPAC), exceptPeptostreptococcus anaerobius(64.6% penicillin-susceptible), remained susceptible to penicillin. AllClostridium perfringensisolates tested were penicillin, metronidazole and meropenem susceptible. OfB. fragilisisolates tested, 5% and 0.83% were meropenem and metronidazole non-susceptible, respectively. Clindamycin susceptibility in anaerobes other than the GPAC is approximately 75% and therefore should not be used as empirical treatment in the absence of susceptibility testing. Considering the global trend of antibiotic resistance among anaerobic bacteria, routine susceptibility testing of anaerobic bacteria, particularly when isolated from critical sites, as well as surveillance of local resistance trends is strongly encouraged. Gradient diffusion MIC determination of anaerobic bacteria is feasible in a clinical diagnostic laboratory and should be more widely utilised.

scholarly journals 2149. Performance of a Gradient Diffusion Method (Etest®) on Mueller–Hinton Agar with Sheep Blood for Aerococcus urinae Antimicrobial Susceptibility Testing

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S728-S729
Author(s):  
Tammy Berteau ◽  
France Emilie Roy ◽  
Julie Bestman-Smith ◽  
Simon Grandjean-Lapierre ◽  
Jean Longtin ◽  
...  
Keyword(s):  
Urinary Tract ◽  
Susceptibility Testing ◽  
Reference Method ◽  
Diffusion Method ◽  
Quebec City ◽  
Sheep Blood ◽  
Maldi Tof ◽  
Gradient Diffusion ◽  
Aerococcus Urinae ◽  
Agar Dilution

Abstract Background Aerococcus urinae is frequently identified by MALDI-TOF in urinary specimens. It is generally susceptible to β-lactams, but its susceptibility pattern to fluoroquinolones (FQ) remains unpredictable. The goal of this study was to evaluate the performance of the gradient diffusion method (Etest®) to determine FQ susceptibility compared with broth microdilution (BMD) and agar dilution (AD). Methods Prospectively collected isolates of A. urinae from urinary tract specimens originating from 5 hospitals in Quebec city and Montreal were identified by MALDI-TOF (Vitek-MS). All isolates were tested using BMD according to CLSI guidelines, and also with Etest® strips on MH agar w/ 5% sheep blood. Isolates showing trailing, insufficient growth or discordance between both methods were further tested by agar dilution (MH agar w/5% horse blood + β-NAD) according to EUCAST guidelines. Breakpoints were interpreted using CLSI M45-A3. Combined results of BMD and AD were then compared with Etest. Results Of the 207 isolates of A. urinae tested, 37 showed trailing (17,8%) and 19 (9,2%) insufficient growth with the BMD method and were retested using AD. Moreover, 38 isolates (ciprofloxacin) and 13 isolates (levofloxacin) showed either lack of categorical or essential agreement between Etest and BMD and were also retested using AD to arbitrate discrepancies. Susceptibility profiles combining BMD and AD are presented in Table 1. As suggested in EUCAST guidelines, readings were much clearer and growth was better with AD compared with BMD. The categorical agreement of the Etest® with BMD+AD was 95% for ciprofloxacin and 97% for levofloxacin. Essential agreement was 95% for ciprofloxacin and 97% for levofloxacin. No very major errors were identified. Two major errors were identified for levofloxacin (1,2%) and one for ciprofloxacin (0.6%). Conclusion Gradient diffusion method using Etest® strips on MH agar w/ sheep blood is a valid method to determine susceptibility to FQ for urinary tract isolates. As a reference method, AD provides clearer endpoints and better growth than BMD for FQ susceptibility testing. Disclosures All authors: No reported disclosures.

scholarly journals Multicenter Clinical Evaluation Of For Susceptibility Testing Of Enterobacterales

2021 ◽  
Author(s):  
Laurine S. Blanchard ◽  
Alex Van Belkum ◽  
Dominique Dechaume ◽  
Tom Armstrong ◽  
Christopher L. Emery ◽  
...  
Keyword(s):  
Susceptibility Testing ◽  
Aminoglycoside Antibiotic ◽  
International Standards ◽  
Diffusion Method ◽  
Individual Species ◽  
Brand Name ◽  
Gradient Diffusion ◽  
The Us ◽  
Tract Infections ◽  
International Standards Organization

Plazomicin (PLZ), brand name ZEMDRI (Cipla Therapeutics), is a novel aminoglycoside antibiotic approved by the US Food and Drug Administration (FDA) for treatment of complicated urinary tract infections including pyelonephritis. ETEST® is a gradient diffusion method that represents an alternative to the more laborious broth micro-dilution (BMD) method for performing antimicrobial susceptibility testing (AST). A multi-center evaluation of the performance of the new ETEST® PLZ (bioMérieux) was conducted in comparison with BMD following FDA and International Standards Organization (ISO) recommendations using FDA-defined breakpoints. Clinical isolates of Enterobacterales (n=598) were included. Fifty-three isolates were resistant to PLZ according to BMD. Overall, the ETEST® PLZ demonstrated 99.0% Essential Agreement (EA), 92.8% Category Agreement (CA), 1.9% Very Major Errors (VME), 0% Major Errors (ME) and 7.0% minor Errors (mE) with both clinical and challenge isolates of Enterobacterales. The VME was found for a single Serratia marcescens strain. Individual species demonstrated EA rates ≥ 90%. In conclusion, we report that ETEST® PLZ represents an accurate tool for performing PLZ AST of Enterobacterales.

scholarly journals ANTIBIOGRAM PROFILE AND BIOFILM FORMING POTENTIAL OF PSEUDOMONAS SPECIES ISOLATED FROM VARIOUS CLINICAL SPECIMENS

2018 ◽  
Vol 11 (10) ◽  
pp. 150
Author(s):  
Jamsheera Cp ◽  
Ethel Suman
Keyword(s):  
Biofilm Formation ◽  
Susceptibility Testing ◽  
Diffusion Method ◽  
Resistance Pattern ◽  
Chi Square ◽  
Clinical Specimens ◽  
Medical College ◽  
Biofilm Production ◽  
Pseudomonas Species ◽  
Chi Square Test

Objective: The present study aimed at finding the resistance pattern of Pseudomonas aeruginosa and other Pseudomonas species isolated from various clinical specimens in the laboratory.Methods: A total of 150 isolates of different species of Pseudomonas obtained from various clinical specimens processed at the Microbiology laboratory of Kasturba Medical College, Manipal Academy of Higher Education, were taken for this study. Antibiotic susceptibility testing was performed by Kirby-Bauer disc diffusion method and interpreted according to the CLSI guidelines. Biofilm assay was performed by modified O’Toole and Kolter method. The results were analyzed using SPSS 17.0 and Student’s unpaired t-test, Kruskal–Wallis, Mann–Whitney, ANOVA, and Chi-square test. p<0.05 was considered statistically significant.Results: Increased resistance was observed by P. aeruginosa to cefotaxime, cotrimoxazole, levofloxacin, ofloxacin, and ticarcillin clavulanate. There was also a good correlation between antibiotic resistance to aztreonam, netilmicin, and ceftazidime and biofilm production. Results of the present study, therefore, demonstrated the occurrence of resistance to various antipseudomonal agents among the biofilm-producing P. aeruginosa isolates.Conclusion: The present study may help in assessing the seriousness of drug resistance caused by biofilm formation in P. aeruginosa and devise strategies through antibiotic policies to minimize such problems.

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