Comparison of BDPhoenix and VITEK2 automated antimicrobial susceptibility test systems for extended-spectrum beta-lactamase detection in Escherichia coli and Klebsiella species clinical isolates.

The present study compares the ability to detect extended-spectrum beta-lactamases (ESBL) among a collection of 34 ESBL producing clinical isolates belonging to Escherichia coli and Klebsiella species with two new rapid susceptibility and identification instruments-VITEK2 (bioMérieux, Marcy l'Etoile, France) vs. BDPhoenix (BD Biosciences, Sparks, MD). ESBL content in these isolates was previously characterized on the basis of PCR amplification and sequencing results which were used as the reference method in our evaluation. BDPhoenix correctly determined the ESBL outcome for all strains tested (100% detection rate), whereas VITEK2 was not able to detect the ESBL status in 5 isolates (85% detection rate). Detailed analysis revealed that the discrepancies were mainly observed with 'difficult-to-detect' strains. Misidentification was either due to low oximino cephalosporin MIC in these strains or was associated with pronounced 'cefotaximase' or 'ceftazidimase' phenotypes. Klebsiella oxytoca chromosomal beta-lactamase (K1) is phenotypically quite similar to ESBL enzymes. In order to evaluate whether the K1 and ESBL enzymes could be discriminated, we expanded our analysis by 8 clinical K. oxytoca strains with K1 phenotypes. VITEK2 gave excellent identification of these strains whereas 7 out of 8 were falsely labeled ESBL-positive by the BDPhoenix system.

[1]  E. S. Moland,et al.  Use of Microdilution Panels with and without β-Lactamase Inhibitors as a Phenotypic Test for β-Lactamase Production among Escherichia coli, Klebsiella spp.,Enterobacter spp., Citrobacter freundii, andSerratia marcescens , 1999, Antimicrobial Agents and Chemotherapy.

[2]  Ellen Jo Baron,et al.  Manual of clinical microbiology , 1975 .

[3]  Mary Jane Ferraro,et al.  Performance standards for antimicrobial susceptibility testing : twelfth informational supplement , 2002 .

[4]  P. Bradford Extended-Spectrum β-Lactamases in the 21st Century: Characterization, Epidemiology, and Detection of This Important Resistance Threat , 2001, Clinical Microbiology Reviews.

[5]  A. Oliver,et al.  Validation of the VITEK2 and the Advance Expert System with a collection of Enterobacteriaceae harboring extended spectrum or inhibitor resistant beta-lactamases. , 2001, Diagnostic microbiology and infectious disease.

[6]  P. Berche,et al.  A Novel Extended-Spectrum TEM-Type β-Lactamase (TEM-52) Associated with Decreased Susceptibility to Moxalactam inKlebsiella pneumoniae , 1998, Antimicrobial Agents and Chemotherapy.

[7]  J. Sutcliffe,et al.  Nucleotide sequence of the ampicillin resistance gene of Escherichia coli plasmid pBR322. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[8]  M. Ferraro,et al.  Antimicrobial susceptibility testing: special needs for fastidious organisms and difficult-to-detect resistance mechanisms. , 2000, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[9]  M. Ferraro,et al.  Antimicrobial susceptibility testing: general principles and contemporary practices. , 1998, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[10]  A P Gibb,et al.  Cefpodoxime screening of Escherichia coli and Klebsiella spp. by Vitek for detection of organisms producing extended-spectrum beta-lactamases. , 2000, Diagnostic microbiology and infectious disease.

[11]  Mary Jane Ferraro,et al.  Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically : approved standard , 2000 .

[12]  Kenneth S. Thomson,et al.  Ability of the VITEK 2 Advanced Expert System To Identify β-Lactam Phenotypes in Isolates of Enterobacteriaceae andPseudomonas aeruginosa , 2000, Journal of Clinical Microbiology.

[13]  Kenneth S. Thomson,et al.  Can Results Obtained with Commercially Available MicroScan Microdilution Panels Serve as an Indicator of β-Lactamase Production among Escherichia coli andKlebsiella Isolates with Hidden Resistance to ExpandedSpectrum Cephalosporins and Aztreonam? , 1998, Journal of Clinical Microbiology.

[14]  I. Stemplinger,et al.  Sequences of beta-lactamase genes encoding CTX-M-1 (MEN-1) and CTX-M-2 and relationship of their amino acid sequences with those of other beta-lactamases , 1996, Antimicrobial agents and chemotherapy.

[15]  D. Livermore beta-Lactamases in laboratory and clinical resistance , 1995, Clinical microbiology reviews.

[16]  E. Kim,et al.  Bloodstream Infections by Extended-Spectrum β-Lactamase-Producing Escherichia coli and Klebsiella pneumoniae in Children: Epidemiology and Clinical Outcome , 2002, Antimicrobial Agents and Chemotherapy.

[17]  A L Barry,et al.  Detection of extended-spectrum-beta-lactamase-producing members of the family Enterobacteriaceae with Vitek ESBL test , 1996, Journal of clinical microbiology.

[18]  Victor L. Yu,et al.  Outcome of Cephalosporin Treatment for Serious Infections Due to Apparently Susceptible Organisms Producing Extended-Spectrum β-Lactamases: Implications for the Clinical Microbiology Laboratory , 2001, Journal of Clinical Microbiology.

[19]  P. Bradford Automated Thermal Cycling Is Superior to Traditional Methods for Nucleotide Sequencing ofblaSHV Genes , 1999, Antimicrobial Agents and Chemotherapy.