scholarly journals Carbapenem Susceptibility Testing Errors Using Three Automated Systems, Disk Diffusion, Etest, and Broth Microdilution and Carbapenem Resistance Genes in Isolates of Acinetobacter baumannii-calcoaceticus Complex

2011 ◽  
Vol 55 (10) ◽  
pp. 4707-4711 ◽  
Author(s):  
Ana Elizabeth Markelz ◽  
Katrin Mende ◽  
Clinton K. Murray ◽  
Xin Yu ◽  
Wendy C. Zera ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Resistance Genes ◽  
Dna Sequences ◽  
Susceptibility Testing ◽  
Carbapenem Resistance ◽  
Disk Diffusion ◽  
Resistance Testing ◽  
Broth Microdilution ◽  
Carbapenemase Gene ◽  
Automated Methods

ABSTRACTTheAcinetobacter baumannii-calcoaceticuscomplex (ABC) is associated with increasing carbapenem resistance, necessitating accurate resistance testing to maximize therapeutic options. We determined the accuracy of carbapenem antimicrobial susceptibility tests for ABC isolates and surveyed them for genetic determinants of carbapenem resistance. A total of 107 single-patient ABC isolates from blood and wound infections from 2006 to 2008 were evaluated. MICs of imipenem, meropenem, and doripenem determined by broth microdilution (BMD) were compared to results obtained by disk diffusion, Etest, and automated methods (the MicroScan, Phoenix, and Vitek 2 systems). Discordant results were categorized as very major errors (VME), major errors (ME), and minor errors (mE). DNA sequences encoding OXA beta-lactamase enzymes (blaOXA-23-like,blaOXA-24-like,blaOXA-58-like, andblaOXA-51-like) and metallo-β-lactamases (MBLs) (IMP, VIM, and SIM1) were identified by PCR, as was theKPC2carbapenemase gene. Imipenem was more active than meropenem and doripenem. The percentage of susceptibility was 37.4% for imipenem, 35.5% for meropenem, and 3.7% for doripenem. Manual methods were more accurate than automated methods.blaOXA-23-likeandblaOXA-24-likewere the primary resistance genes found.blaOXA-58-like, MBLs, andKPC2were not present. Both automated testing and manual testing for susceptibility to doripenem were very inaccurate, with VME rates ranging between 2.8 and 30.8%. International variability in carbapenem breakpoints and the absence of CLSI breakpoints for doripenem present a challenge in susceptibility testing.

scholarly journals Horizontal Transfer of the OXA-24 Carbapenemase Gene via Outer Membrane Vesicles: a New Mechanism of Dissemination of Carbapenem Resistance Genes in Acinetobacter baumannii

2011 ◽  
Vol 55 (7) ◽  
pp. 3084-3090 ◽  
Author(s):  
Carlos Rumbo ◽  
Esteban Fernández-Moreira ◽  
María Merino ◽  
Margarita Poza ◽  
Jose Antonio Mendez ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Outer Membrane ◽  
Resistance Genes ◽  
Carbapenem Resistance ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Content Type ◽  
Class D ◽  
Carbapenemase Gene

ABSTRACTThe resistance ofAcinetobacter baumanniistrains to carbapenems is a worrying problem in hospital settings. The main mechanism of carbapenem resistance is the expression of β-lactamases (metalloenzymes or class D enzymes). The mechanisms of the dissemination of these genes amongA. baumanniistrains are not fully understood. In this study we used two carbapenem-resistant clinical strains ofA. baumannii(AbH12O-A2 and AbH12O-CU3) expressing the plasmid-borneblaOXA-24gene (plasmids pMMA2 and pMMCU3, respectively) to demonstrate thatA. baumanniireleases outer membrane vesicles (OMVs) duringin vitrogrowth. The use of hybridization studies enabled us to show that these OMVs harbored theblaOXA-24gene. The incubation of these OMVs with the carbapenem-susceptibleA. baumanniiATCC 17978 host strain yielded full resistance to carbapenems. The presence of the original plasmids harboring theblaOXA-24gene was detected in strain ATCC 17978 after the transformation of OMVs. New OMVs harboringblaOXA-24were released byA. baumanniiATCC 17978 after it was transformed with the original OMV-mediated plasmids, indicating the universality of the process. We present the first experimental evidence that clinical isolates ofA. baumanniimay release OMVs as a mechanism of horizontal gene transfer whereby carbapenem resistance genes are delivered to surroundingA. baumanniibacterial isolates.

scholarly journals Pseudo-Outbreak of Imipenem-Resistant Acinetobacter baumannii Resulting from False Susceptibility Testing by a Rapid Automated System

2000 ◽  
Vol 38 (9) ◽  
pp. 3505-3507 ◽  
Author(s):  
Athanassios Tsakris ◽  
Anastassia Pantazi ◽  
Spyros Pournaras ◽  
Antonios Maniatis ◽  
Aleka Polyzou ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
General Hospital ◽  
Susceptibility Testing ◽  
Homogeneous Group ◽  
Susceptibility Test ◽  
Automated System ◽  
Disk Diffusion ◽  
Broth Microdilution ◽  
High Prevalence ◽  
Agar Dilution

Introduction of the Vitek GNS-506 susceptibility testing cards in the Hippokration General Hospital, Thessaloniki, Greece, resulted in an apparently high prevalence of imipenem-resistant Acinetobacter baumannii. When 35 of these isolates were further tested by disk diffusion, broth microdilution, and agar dilution assays, 32 were imipenem sensitive by all tests and three were sensitive or intermediate, depending on the method. The pseudoresistant acinetobacters did not form a genetically homogeneous group. It is suggested that the detection of imipenem-resistant A. baumannii isolates by this system should be confirmed by an additional susceptibility test.

scholarly journals Comparative Evaluation of the VITEK 2, Disk Diffusion, Etest, Broth Microdilution, and Agar Dilution Susceptibility Testing Methods for Colistin in Clinical Isolates, Including Heteroresistant Enterobacter cloacae and Acinetobacter baumannii Strains

2007 ◽  
Vol 51 (10) ◽  
pp. 3726-3730 ◽  
Author(s):  
Jerome R. Lo-Ten-Foe ◽  
Anne Marie G. A. de Smet ◽  
Bram M. W. Diederen ◽  
Jan A. J. W. Kluytmans ◽  
Peter H. J. van Keulen
Keyword(s):  
Acinetobacter Baumannii ◽  
Susceptibility Testing ◽  
Enterobacter Cloacae ◽  
Clinical Isolates ◽  
Disk Diffusion ◽  
Broth Microdilution ◽  
Diffusion Test ◽  
Disk Diffusion Test ◽  
Agar Dilution ◽  
Vitek 2

ABSTRACT Increasing antibiotic resistance in gram-negative bacteria has recently renewed interest in colistin as a therapeutic option. The increasing use of colistin necessitates the availability of rapid and reliable methods for colistin susceptibility testing. We compared seven methods of colistin susceptibility testing (disk diffusion, agar dilution on Mueller-Hinton [MH] and Isosensitest agar, Etest on MH and Isosensitest agar, broth microdilution, and VITEK 2) on 102 clinical isolates collected from patient materials during a selective digestive decontamination or selective oral decontamination trial in an intensive-care unit. Disk diffusion is an unreliable method to measure susceptibility to colistin. High error rates and low levels of reproducibility were observed in the disk diffusion test. The colistin Etest, agar dilution, and the VITEK 2 showed a high level of agreement with the broth microdilution reference method. Heteroresistance for colistin was observed in six Enterobacter cloacae isolates and in one Acinetobacter baumannii isolate. This is the first report of heteroresistance to colistin in E. cloacae isolates. Resistance to colistin in these isolates seemed to be induced upon exposure to colistin rather than being caused by stable mutations. Heteroresistant isolates could be detected in the broth microdilution, agar dilution, Etest, or disk diffusion test. The VITEK 2 displayed low sensitivity in the detection of heteroresistant subpopulations of E. cloacae. The VITEK 2 colistin susceptibility test can therefore be considered to be a reliable tool to determine susceptibility to colistin in isolates of genera that are known not to exhibit resistant subpopulations. In isolates of genera known to (occasionally) exhibit heteroresistance, an alternative susceptibility testing method capable of detecting heteroresistance should be used.

scholarly journals Comparison of Minocycline Susceptibility Testing Methods for Carbapenem-Resistant Acinetobacter baumannii

2016 ◽  
Vol 54 (12) ◽  
pp. 2937-2941 ◽  
Author(s):  
Peng Wang ◽  
Sarah L. Bowler ◽  
Serena F. Kantz ◽  
Roberta T. Mettus ◽  
Yan Guo ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Susceptibility Testing ◽  
Error Rates ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Broth Microdilution ◽  
Testing Methods ◽  
Content Type ◽  
Minor Error ◽  
Carbapenem Resistant

Treatment options for infections due to carbapenem-resistantAcinetobacter baumanniiare extremely limited. Minocycline is a semisynthetic tetracycline derivative with activity against this pathogen. This study compared susceptibility testing methods that are used in clinical microbiology laboratories (Etest, disk diffusion, and Sensititre broth microdilution methods) for testing of minocycline, tigecycline, and doxycycline against 107 carbapenem-resistantA. baumanniiclinical isolates. Susceptibility rates determined with the standard broth microdilution method using cation-adjusted Mueller-Hinton (MH) broth were 77.6% for minocycline and 29% for doxycycline, and 92.5% of isolates had tigecycline MICs of ≤2 μg/ml. Using MH agar from BD and Oxoid, susceptibility rates determined with the Etest method were 67.3% and 52.3% for minocycline, 21.5% and 18.7% for doxycycline, and 71% and 29.9% for tigecycline, respectively. With the disk diffusion method using MH agar from BD and Oxoid, susceptibility rates were 82.2% and 72.9% for minocycline and 34.6% and 34.6% for doxycycline, respectively, and rates of MICs of ≤2 μg/ml were 46.7% and 23.4% for tigecycline. In comparison with the standard broth microdilution results, very major rates were low (∼2.8%) for all three drugs across the methods, but major error rates were higher (∼5.6%), especially with the Etest method. For minocycline, minor error rates ranged from 14% to 37.4%. For tigecycline, minor error rates ranged from 6.5% to 69.2%. The majority of minor errors were due to susceptible results being reported as intermediate. For minocycline susceptibility testing of carbapenem-resistantA. baumanniistrains, very major errors are rare, but major and minor errors overcalling strains as intermediate or resistant occur frequently with susceptibility testing methods that are feasible in clinical laboratories.

scholarly journals WGS based analysis of acquired antimicrobial resistance in human and non-human Acinetobacter baumannii isolates from a German perspective

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Gamal Wareth ◽  
Christian Brandt ◽  
Lisa D. Sprague ◽  
Heinrich Neubauer ◽  
Mathias W. Pletz
Keyword(s):  
Antimicrobial Resistance ◽  
Acinetobacter Baumannii ◽  
Genome Sequencing ◽  
Resistance Genes ◽  
In Silico ◽  
Acquired Resistance ◽  
Whole Genome ◽  
Chromosomal Gene ◽  
Milk Samples ◽  
Carbapenemase Gene

Abstract Background Acinetobacter baumannii ability to develop and acquire resistance makes it one of the most critical nosocomial pathogens globally. Whole-genome sequencing (WGS) was applied to identify the acquired or mutational variants of antimicrobial resistance (AMR) genes in 85 German A. baumannii strains utilizing Illumina technology. Additionally, the whole genome of 104 German isolates deposited in the NCBI database was investigated. Results In-silico analysis of WGS data revealed wide varieties of acquired AMR genes mediating resistance mostly to aminoglycosides, cephalosporins, carbapenems, sulfonamides, tetracyclines and macrolides. In the 189 analyzed genomes, the ant (3″)-IIa conferring resistance to aminoglycosides was the most frequent (55%), followed by blaADC.25 (38.6%) conferring resistance to cephalosporin, blaOXA-23 (29%) and the blaOXA-66 variant of the intrinsic blaOXA-51-likes (26.5%) conferring resistance to carbapenems, the sul2 (26%) conferring resistance to sulfonamides, the tet. B (19.5%) conferring resistance to tetracycline, and mph. E and msr. E (19%) conferring resistance to macrolides. blaTEM variants conferring resistance to cephalosporins were found in 12% of genomes. Thirteen variants of the intrinsic blaOXA-51 carbapenemase gene, blaOXA-510 and blaADC-25 genes were found in isolates obtained from dried milk samples. Conclusion The presence of strains harboring acquired AMR genes in dried milk raises safety concerns and highlights the need for changes in producing dried milk. Acquired resistance genes and chromosomal gene mutation are successful routes for disseminating AMR determinants among A. baumannii. Identification of chromosomal and plasmid-encoded AMR in the genome of A. baumannii may help understand the mechanism behind the genetic mobilization and spread of AMR genes.

scholarly journals Tetracycline Susceptibility Testing and Resistance Genes in Isolates of Acinetobacter baumannii-Acinetobacter calcoaceticus Complex from a U.S. Military Hospital

2009 ◽  
Vol 53 (6) ◽  
pp. 2693-2695 ◽  
Author(s):  
Kevin S. Akers ◽  
Katrin Mende ◽  
Heather C. Yun ◽  
Duane R. Hospenthal ◽  
Miriam L. Beckius ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Resistance Genes ◽  
Susceptibility Testing ◽  
Tetracycline Resistance ◽  
Acinetobacter Calcoaceticus ◽  
Multidrug Resistant ◽  
Military Hospital ◽  
Tetracycline Resistance Genes ◽  
Testing Method

ABSTRACT Infections with multidrug-resistant Acinetobacter baumannii-Acinetobacter calcoaceticus complex bacteria complicate the care of U.S. military personnel and civilians worldwide. One hundred thirty-three isolates from 89 patients at our facility during 2006 and 2007 were tested by disk diffusion, Etest, and broth microdilution for susceptibility to tetracycline, doxycycline, minocycline, and tigecycline. Minocycline was the most active in vitro, with 90% of the isolates tested susceptible. Susceptibilities varied significantly with the testing method. The acquired tetracycline resistance genes tetA, tetB, and tetA(39) were present in the isolates.

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