scholarly journals Performance Evaluation of the Gradient Diffusion Strip Method and Disk Diffusion Method for Ceftazidime–Avibactam Against Enterobacterales and Pseudomonas aeruginosa: A Dual-Center Study

2021 ◽  
Vol 12 ◽  
Author(s):  
Jingjia Zhang ◽  
Gang Li ◽  
Ge Zhang ◽  
Wei Kang ◽  
Simeng Duan ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Carbapenem Resistance ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Major Error ◽  
Beta Lactam ◽  
Disk Diffusion Method ◽  
Strip Method ◽  
Gradient Diffusion

Objectives: Ceftazidime–avibactam is a novel synthetic beta-lactam + beta-lactamase inhibitor combination. We evaluated the performance of the gradient diffusion strip method and the disk diffusion method for the determination of ceftazidime–avibactam against Enterobacterales and Pseudomonas aeruginosa.Methods: Antimicrobial susceptibility testing of 302 clinical Enterobacterales and Pseudomonas aeruginosa isolates from two centers were conducted by broth microdilution (BMD), gradient diffusion strip method, and disk diffusion method for ceftazidime–avibactam. Using BMD as a gold standard, essential agreement (EA), categorical agreement (CA), major error (ME), and very major error (VME) were determined according to CLSI guidelines. CA and EA rate > 90%, ME rate < 3%, and VME rate < 1.5% were considered as acceptable criteria. Polymerase chain reaction and Sanger sequencing were performed to determine the carbapenem resistance genes of all 302 isolates.Results: A total of 302 strains were enrolled, among which 182 strains were from center 1 and 120 strains were from center 2. A percentage of 18.21% (55/302) of the enrolled isolates were resistant to ceftazidime–avibactam. The CA rates of the gradient diffusion strip method for Enterobacterales and P. aeruginosa were 100% and 98.65% (73/74), respectively, and the EA rates were 97.37% (222/228) and 98.65% (73/74), respectively. The CA rates of the disk diffusion method for Enterobacterales and P. aeruginosa were 100% and 95.95% (71/74), respectively. No VMEs were found by using the gradient diffusion strip method, while the ME rate was 0.40% (1/247). No MEs were found by using the disk diffusion method, but the VME rate was 5.45% (3/55). Therefore, all the parameters of the gradient diffusion strip method were in line with acceptable criteria. For 31 blaKPC, 33 blaNDM, 7 blaIMP, and 2 blaVIM positive isolates, both CA and EA rates were 100%; no MEs or VMEs were detected by either method. For 15 carbapenemase-non-producing resistant isolates, the CA and EA rates of the gradient diffusion strips method were 100%. Whereas the CA rate of the disk diffusion method was 80.00% (12/15), the VME rate was 20.00% (3/15).Conclusion: The gradient diffusion strip method can meet the needs of clinical microbiological laboratories for testing the susceptibility of ceftazidime–avibactam drugs. However, the VME rate > 1.5% (5.45%) by the disk diffusion method. By comparison, the performance of the gradient diffusion strip method was better than that of the disk diffusion method.

scholarly journals Presence of different beta-lactamase classes among clinical isolates of Pseudomonas aeruginosa expressing AmpC beta-lactamase enzyme

2010 ◽  
Vol 4 (04) ◽  
pp. 239-242 ◽  
Author(s):  
Supriya Upadhyay ◽  
Malay Ranjan Sen ◽  
Amitabha Bhattacharjee
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Treatment Options ◽  
Clinical Isolates ◽  
Diffusion Method ◽  
Beta Lactamase ◽  
Beta Lactam ◽  
Disk Diffusion Method ◽  
Extended Spectrum Beta Lactamase ◽  
Extended Spectrum ◽  
Enzyme Detection

Introduction: Infections caused by Pseudomonas aeruginosa are difficult to treat as the majority of isolates exhibit varying degrees of beta-lactamase mediated resistance to most of the beta-lactam antibiotics. It is also not unusual to find a single isolate that expresses multiple β-lactamase enzymes, further complicating the treatment options. Thus the present study was designed to investigate the coexistence of different beta-lactamase enzymes in clinical isolates of P. aeruginosa. Methodology: A total of 202 clinical isolates of P. aeruginosa were tested for the presence of AmpC beta-lactamase, extended spectrum beta-lactamase (ESBL) and metallo beta-lactamase (MBL) enzyme. Detection of AmpC beta-lactamase was performed by disk antagonism test and a modified three-dimensional method, whereas detection of ESBL was done by the combined disk diffusion method per Clinical and Laboratory Standards Institute (CLSI) guidelines and MBL were detected by the Imipenem EDTA disk potentiation test. Results: A total of 120 (59.4%) isolates were confirmed to be positive for AmpC beta-lactamase. Among them, 14 strains (7%) were inducible AmpC producers. Co-production of AmpC along with extended spectrum beta-lactamase and metallo beta-lactamase was reported in 3.3% and 46.6% isolates respectively. Conclusion: The study emphasizes the high prevalence of multidrug resistant P. aeruginosa producing beta-lactamase enzymes of diverse mechanisms. Thus proper antibiotic policy and measures to restrict the indiscriminative use of cephalosporins and carbapenems should be taken to minimize the emergence of this multiple beta-lactamase producing pathogens.

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.

RELIABILITY OF A DISK DIFFUSION METHOD USING SEMICONFLUENT GROWTH IN THE DETERMINATION OF AMINOGLYCOSIDE RESISTANCE

2009 ◽  
Vol 94B (1-6) ◽  
pp. 153-157
Author(s):  
PENTTI Huovinen ◽  
ELJA Herva ◽  
MARJA-LEENA Katila ◽  
MARJA-LIISA Klossner ◽  
OLLI-VEIKKO Renkonen ◽  
...  
Keyword(s):  
Disk Diffusion ◽  
Diffusion Method ◽  
Aminoglycoside Resistance ◽  
Disk Diffusion Method

scholarly journals Occurrence of NDM-1 and VIM-2 Co-Producing Escherichia coli and OprD Alteration in Pseudomonas aeruginosa Isolated from Hospital Environment Samples in Northwestern Tunisia

Diagnostics ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 1617
Author(s):  
Raouaa Maaroufi ◽  
Olfa Dziri ◽  
Linda Hadjadj ◽  
Seydina M. Diene ◽  
Jean-Marc Rolain ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Bacterial Resistance ◽  
Carbapenem Resistance ◽  
Multidrug Resistant ◽  
Hospital Environment ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Multidrug Resistant Bacteria ◽  
Disk Diffusion Method

Hospital environments constitute the main reservoir of multidrug-resistant bacteria. In this study we aimed to investigate the presence of Gram-negative bacteria in one Northwestern Tunisian hospital environment, and characterize the genes involved in bacterial resistance. A total of 152 environmental isolates were collected from various surfaces and isolated using MacConkey medium supplemented with cefotaxime or imipenem, with 81 fermenter bacteria (27 Escherichia coli, and 54 Enterobacter spp., including 46 Enterobacter cloacae), and 71 non-fermenting bacteria (69 Pseudomonas spp., including 54 Pseudomonas aeruginosa, and 2 Stenotrophomonas maltophilia) being identified by the MALDI-TOF-MS method. Antibiotic susceptibility testing was performed by disk diffusion method and E-Test was used to determine MICs for imipenem. Several genes implicated in beta-lactams resistance were characterized by PCR and sequencing. Carbapenem resistance was detected among 12 isolates; nine E. coli (blaNDM-1 (n = 8); blaNDM-1 + blaVIM-2 (n = 1)) and three P. aeruginosa were carbapenem-resistant by loss of OprD porin. The whole-genome sequencing of P. aeruginosa 97H was determined using Illumina MiSeq sequencer, typed ST285, and harbored blaOXA-494. Other genes were also detected, notably blaTEM (n = 23), blaCTX-M-1 (n = 10) and blaCTX-M-9 (n = 6). These new epidemiological data imposed new surveillance strategies and strict hygiene rules to decrease the spread of multidrug-resistant bacteria in this area.

scholarly journals In vitro efficacy of synergistic antibiotic combinations in imipenem resistant Pseudomonas aeruginosa strains

2017 ◽  
Vol 9 (1) ◽  
pp. 3-8
Author(s):  
Aleya Farzana ◽  
S. M. Shamsuzzaman
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Carbapenem Resistance ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Considerable Proportion ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Disk Diffusion Method ◽  
New Antibiotics

The increase in antibiotic resistance coincided with the decline in production of new antibiotics. Combination antibiotic treatment is preferred in nosocomial infections caused by multidrug resistant Pseudomonas aeruginosa. In vitro synergism test by agar dilution method were used to choose the combinations which might be used in clinic. The aim of this study was to investigate the synergistic efficacy of antibiotic combinations in imipenem resistant P. aeruginosa strains. Carbapenem resistance (imipenem and meropenem) wasdetermined by disk diffusion method. Among isolated P. aeruginosa 44.9% were cabapenem resistant. The MIC of drugs among 25 imipenem resistant isolates ranged from >_ 256 mg/L to <_ 8 mg/L for imipenem, >_ 1024 mg/L to <_ 64 mg/L for ceftriaxone, >_ 256 mg/L to <_ 8 mg/L for amikacin, >_ 16 mg/L to <_ 2 mg/L for colistin, >_ 512 mg/L to <_ 16 mg/L for piperacillin/tazobactam. Among antibiotic combinations, piperacillin /tazobactam- amikacin was most effective with 80% synergism next to which was imipenem-amikacin with 60% synergism, then imipenem-colistin with 50% synergism, imipenem-ceftriaxone with 30% synergism. Only one combination (piperacillin/tazobactum -imipenem showed 20% antagonism. All these combinations had considerable proportion of additive effect which is also desirable for these multi drug resistant isolates.Bangladesh J Med Microbiol 2015; 9 (1): 3-8

scholarly journals Assay of antibiotic detection limits in cow’s milk model samples and comparison of sensitivity of various detection systems (disk diffusion method, Delvotest SP and Penzym S 100).

2013 ◽  
Vol 19 (No. 4) ◽  
pp. 125-131 ◽  
Author(s):  
B. Hozová ◽  
M. Kratmüllerová
Keyword(s):  
Bacillus Stearothermophilus ◽  
Detection Limits ◽  
High Sensitivity ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Beta Lactam ◽  
Disk Diffusion Method ◽  
Detection Systems ◽  
Beta Lactam Antibiotics ◽  
S 100

The work was aimed at estimating the detection limits of 21 antibiotics (beta-lactam antibiotics, cephalosporins, aminoglycosides, macrolides and others) by means of three types of detection systems (disk diffusion method with&nbsp;Bacillus stearothermophilus var. calidolactis C 953, Delvotest SP and Penzym S 100) and to verify their sensitivity in the evaluation of the admissible maximum residual limits (MRL). The high sensitivity and the good correlation of results have been achieved mainly by applying the rapid methods such as Delvotest SP and Penzym S 100 versus the less sensitive disk diffusion method.&nbsp; In the next stage of the work, detection limits of the mutual combinations of antibiotics in milk were estimated. In the model experiment, the synergic effect between ampicillin and oxacillin, cefuroxime and trimethoprim and between cephalosporins (combination of cefazolin with&nbsp;cefoperhazon) was observed.

scholarly journals Escherichia coli Overexpressing a Baeyer-Villiger Monooxygenase from Acinetobacter radioresistens Becomes Resistant to Imipenem

2015 ◽  
Vol 60 (1) ◽  
pp. 64-74 ◽  
Author(s):  
Daniela Minerdi ◽  
Ivan Zgrablic ◽  
Silvia Castrignanò ◽  
Gianluca Catucci ◽  
Claudio Medana ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bacterial Infections ◽  
Bacterial Species ◽  
Carbapenem Resistance ◽  
Multidrug Resistant ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Content Type ◽  
Acinetobacter Radioresistens

ABSTRACTAntimicrobial resistance is a global issue currently resulting in the deaths of hundreds of thousands of people a year worldwide. Data present in the literature illustrate the emergence of many bacterial species that display resistance to known antibiotics;Acinetobacterspp. are a good example of this. We report here thatAcinetobacter radioresistenshas a Baeyer-Villiger monooxygenase (Ar-BVMO) with 100% amino acid sequence identity to the ethionamide monooxygenase of multidrug-resistant (MDR)Acinetobacter baumannii. Both enzymes are only distantly phylogenetically related to other canonical bacterial BVMO proteins. Ar-BVMO not only is capable of oxidizing two anticancer drugs metabolized by human FMO3, danusertib and tozasertib, but also can oxidize other synthetic drugs, such as imipenem. The latter is a member of the carbapenems, a clinically important antibiotic family used in the treatment of MDR bacterial infections. Susceptibility tests performed by the Kirby-Bauer disk diffusion method demonstrate that imipenem-sensitiveEscherichia coliBL21 cells overexpressing Ar-BVMO become resistant to this antibiotic. An agar disk diffusion assay proved that when imipenem reacts with Ar-BVMO, it loses its antibiotic property. Moreover, an NADPH consumption assay with the purified Ar-BVMO demonstrates that this antibiotic is indeed a substrate, and its product is identified by liquid chromatography-mass spectrometry to be a Baeyer-Villiger (BV) oxidation product of the carbonyl moiety of the β-lactam ring. This is the first report of an antibiotic-inactivating BVMO enzyme that, while mediating its usual BV oxidation, also operates by an unprecedented mechanism of carbapenem resistance.

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 Etests and the disk diffusion method for assessment of the activity of ceftazidime-avibactam against Enterobacterales and Pseudomonas aeruginosa in China

2020 ◽  
Author(s):  
Qi Wang ◽  
Feifei Zhang ◽  
Zhanwei Wang ◽  
Hongbin Chen ◽  
Xiaojuan Wang ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Random Selection ◽  
Alternative Methods ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Broth Microdilution ◽  
Antimicrobial Susceptibility Test ◽  
Disk Diffusion Method ◽  
Gram Negative ◽  
Selection Group

Abstract Background: Gram-negative bacilli, particularly Enterobacterales and Pseudomonas aeruginosa, often acquire antimicrobial resistance. Ceftazidime-avibactam was approved for use in China in 2019. However, currently available commercial antimicrobial susceptibility test kits have not yet been developed. Here, we evaluated the Etest and disk diffusion method for assessment of the efficacy of ceftazidime-avibactam against Enterobacterales and P. aeruginosa in China. Results: In total, 194 Enterobacterales and 77 P. aeruginosa isolates, which were divided into a random selection group (140 Enterobacterales and 54 P. aeruginosa isolates) and a stock group (46 Enterobacterales and 31 P. aeruginosa isolates), were assessed by the Etest, disk diffusion, and broth microdilution (BMD) methods. Minimum inhibitory concentrations (MICs) and zone diameters were interpreted according to the CLSI M100 30th edition. For all 271 Enterobacterales and P. aeruginosa isolates, no very major errors were found using Etests. The overall categorical agreement rates (CA%) of Etests for Enterobacterales and P. aeruginosa were 99.5% (193/194) and 96.1% (74/77), respectively. The overall essential agreement rates (EA%) of Etests for Enterobacterales and P. aeruginosa were 95.9% (186/194) and 94.8% (73/77), respectively. In both the random selection and stock groups, EA% and CA% values of Etests exceeded 90%. Overall CA% values of the disk diffusion method for Enterobacterales and P. aeruginosa were 98.5% (191/194) and 93.5% (71/77), respectively. There was no linear relationship between zone diameter and BMD MIC. Conclusions: For Enterobacterales and P. aeruginosa, Etests and the disk diffusion method could have better performance as alternative methods to meet the needs of clinical treatment interpretation. Application of the disk diffusion method in Enterobacterales was superior to that in P. aeruginosa.

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