Evaluation of double-disk potentiation and disk potentiation tests using dipicolinic acid for detection of metallo-β-lactamase-producing pseudomonas spp. and Acinetobacter spp

J Clin Microbiol. 2012 Oct;50(10):3227-32. doi: 10.1128/JCM.00818-12. Epub 2012 Jul 25.

Abstract

Accurate detection of metallo-β-lactamase (MBL)-producing Pseudomonas spp. and Acinetobacter spp. became very important with the increasing prevalence of carbapenem-nonsusceptible clinical isolates. The performance of phenotypic MBL detection methods may depend on the types of MBL and the characteristics of the isolates. A high false-positive rate is a problem with EDTA-based MBL detection methods. We evaluated the performance of double-disk potentiation tests (DDPTs) and disk potentiation tests (DPTs) with dipicolinic acid (DPA) using 44 isolates of Pseudomonas spp. and Acinetobacter spp. producing IMP-1-like, VIM-2-like, and SIM-1 type MBLs. Also, we characterized P. aeruginosa isolates with positive imipenem (IPM)-DPA DDPT, but negative meropenem (MEM)-DPA DDPT, and determined possibility of improving a DDPT by using MacConkey agar. Among five different DDPT methods, the IPM-DPA 250-μg method showed the highest sensitivity (97.7%) and specificity (100%). Among four DPT tests, the highest sensitivity (100%) was shown by the IPM-EDTA 1,900-μg disk method, but the specificity was very low (11.4%). Five of six P. aeruginosa isolates with false-negative DDPTs with MEM-DPA 250-μg disks carried bla(IMP-6,) and the high level resistance to MEM (MIC ≥ 512 μg/ml) was reduced by the presence of phenylalanine arginine β-naphtylamide. Improvement of DDPTs was observed when MacConkey agar was used instead of Mueller-Hinton agar. In conclusion, DPA is a better MBL inhibitor than EDTA for detection of Pseudomonas spp. and Acinetobacter spp. with IMP-1-like, VIM-2-like, and SIM-1-type MBLs. In DPA DDPTs, IPM disks perform better than MEM disks when the isolates are highly resistant to MEM due to the overexpression of efflux pumps.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acinetobacter / enzymology*
  • Acinetobacter / isolation & purification
  • Acinetobacter Infections / microbiology
  • Bacteriological Techniques / methods*
  • Culture Media / chemistry
  • Humans
  • Picolinic Acids / metabolism
  • Pseudomonas / enzymology*
  • Pseudomonas / isolation & purification
  • Pseudomonas Infections / microbiology
  • Sensitivity and Specificity
  • beta-Lactamases / analysis*

Substances

  • Culture Media
  • Picolinic Acids
  • beta-Lactamases
  • dipicolinic acid