Broth microdilution methods using B-lactamase inhibitors for the identification of Klebsiella pneumoniae carbapenemases and metallo-B-lactamases in gram-negative bacilli

Seokhoon Jeong, Wonkeun Song, Il Kwon Bae, Han Sung Kim, Jae Seok Kim, Min Jeong Park, Dong Hoon Shin

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

This study was designed to evaluate the performance of the broth microdilution (BMD) test in detecting the production of Klebsiella pneumoniae carbapenemases (KPCs) and metallo-β-lactamases(MBLs) in clinical isolates of gram-negative bacilli by using various β-lactamase inhibitors. A carbapenemase detection test, comprised of Mueller-Hinton broth containing serial twofold dilutions of imipenem or meropenem with and without aminophenylboronic acid (APB), phenylboronic acid (PB), cloxacillin(CLX), dipicolinic acid (DPA), or EDTA, was evaluated against 31 Klebsiella pneumoniae strains with KPC, 21 MBL-producers (3 Enterobacteriaceae and 18 non-fermenters), and 26 carbapenemase-non-producing Enterobacteriaceae strains. A threefold or greater decrease in the MIC of imipenem or meropenem with β-lactamase inhibitors, when compared with the imipenem or meropenem alone, was considered a positive result. Imipenem with and without PB had the most comparable sensitivity (93.5%) and specificity (93.6%) for the detection of K. pneumoniae with KPC enzymes if the additional criterion of a negative CLX result was included. Both DPA and EDTA had excellent sensitivity (imipenem with and without DPA vs. EDTA, 90.5% vs. 95.2%) and specificity (imipenem with and without DPA vs. EDTA, 98.2% vs. 100%) for the detection of MBL-producing gram-negative bacilli. The comparative study showed that, by using imipenem with and without PB, CLX, and EDTA, the BMD test was effective in detecting KPC and MBL enzymes. The BMD test could be applied for routine use in commercially available semiautomated systems for the detection of KPCs and MBLs in gram-negative bacilli.

Original languageEnglish
Pages (from-to)49-55
Number of pages7
JournalAnnals of Clinical and Laboratory Science
Volume44
Issue number1
Publication statusPublished - 2014 Jan 1

Fingerprint

Imipenem
Klebsiella pneumoniae
Bacilli
meropenem
Bacillus
Edetic Acid
Cloxacillin
Enterobacteriaceae
Enzymes
Dilution
Sensitivity and Specificity
dipicolinic acid
benzeneboronic acid

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology and Allergy
  • Pathology and Forensic Medicine
  • Immunology
  • Molecular Biology
  • Hematology
  • Clinical Biochemistry
  • Medical Laboratory Technology

Cite this

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title = "Broth microdilution methods using B-lactamase inhibitors for the identification of Klebsiella pneumoniae carbapenemases and metallo-B-lactamases in gram-negative bacilli",
abstract = "This study was designed to evaluate the performance of the broth microdilution (BMD) test in detecting the production of Klebsiella pneumoniae carbapenemases (KPCs) and metallo-β-lactamases(MBLs) in clinical isolates of gram-negative bacilli by using various β-lactamase inhibitors. A carbapenemase detection test, comprised of Mueller-Hinton broth containing serial twofold dilutions of imipenem or meropenem with and without aminophenylboronic acid (APB), phenylboronic acid (PB), cloxacillin(CLX), dipicolinic acid (DPA), or EDTA, was evaluated against 31 Klebsiella pneumoniae strains with KPC, 21 MBL-producers (3 Enterobacteriaceae and 18 non-fermenters), and 26 carbapenemase-non-producing Enterobacteriaceae strains. A threefold or greater decrease in the MIC of imipenem or meropenem with β-lactamase inhibitors, when compared with the imipenem or meropenem alone, was considered a positive result. Imipenem with and without PB had the most comparable sensitivity (93.5{\%}) and specificity (93.6{\%}) for the detection of K. pneumoniae with KPC enzymes if the additional criterion of a negative CLX result was included. Both DPA and EDTA had excellent sensitivity (imipenem with and without DPA vs. EDTA, 90.5{\%} vs. 95.2{\%}) and specificity (imipenem with and without DPA vs. EDTA, 98.2{\%} vs. 100{\%}) for the detection of MBL-producing gram-negative bacilli. The comparative study showed that, by using imipenem with and without PB, CLX, and EDTA, the BMD test was effective in detecting KPC and MBL enzymes. The BMD test could be applied for routine use in commercially available semiautomated systems for the detection of KPCs and MBLs in gram-negative bacilli.",
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Broth microdilution methods using B-lactamase inhibitors for the identification of Klebsiella pneumoniae carbapenemases and metallo-B-lactamases in gram-negative bacilli. / Jeong, Seokhoon; Song, Wonkeun; Bae, Il Kwon; Kim, Han Sung; Kim, Jae Seok; Park, Min Jeong; Shin, Dong Hoon.

In: Annals of Clinical and Laboratory Science, Vol. 44, No. 1, 01.01.2014, p. 49-55.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Broth microdilution methods using B-lactamase inhibitors for the identification of Klebsiella pneumoniae carbapenemases and metallo-B-lactamases in gram-negative bacilli

AU - Jeong, Seokhoon

AU - Song, Wonkeun

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AU - Shin, Dong Hoon

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