title: Detection of metallo-β-lactamases producing Acinetobacter baumannii using microbiological assay, disc synergy test and PCR.
abstract: BACKGROUND
  One leading factor responsible for resistance in Acinetobacter baumannii, an important opportunist in health care institutions globally, is the production of carbapenamases like metallo-β-lactamases (MBLs), which hydrolyze a variety of β-lactams including penicillin, cephalosporins and carbapenems. However, neither any standard guidelines are available nor any method has been found to be perfect for their detection. Various methods have shown discordant results, depending upon the employed methodology, β-lactamase substrate and MBL inhibitor used. This study aims to evaluate two phenotypic methods against PCR as gold standard among carbapenem resistant A. baumannii for identifying MBL producers.
  
  
  METHODS
  A total of 130 A. baumannii were screened for imipenem and meropenem resistance by Kirby-Bauer disc diffusion method. Phenotypic expression of MBL was detected by EDTA-imipenem-microbiological (EIM) assay and extended EDTA disc synergy (eEDS) test and presence of bla-IMP and bla-VIM was detected by PCR in all the carbapenem resistant isolates.
  
  
  RESULTS
  Of the 43 imipenem and/or meropenem resistant A. baumannii isolates, 4 (9.3%) were found to be MBL producers by EIM and 3 (6.97%) by eEDS. Only bla-VIM gene was detected in 7 (16.28%) by PCR. In addition EIM detected 14 (32.56%) carbapenem resistant non-metallo enzyme producers.
  
  
  CONCLUSIONS
  Of the two MBL genes targeted, bla-VIM was only detected and that too in isolates resistant to both imipenem and meropenem. Further, EIM was useful in differentiating MBL from non-metalloenzymes producers.
authors.affiliation: Department of Microbiology, Mahatma Gandhi Institute of Medical Sciences, Sevagram, Wardha, India.
authors.forename: M
authors.initials: M
authors.lastname: Purohit
authors.forename: D K
authors.initials: DK
authors.lastname: Mendiratta
authors.forename: V S
authors.initials: VS
authors.lastname: Deotale
authors.forename: M
authors.initials: M
authors.lastname: Madhan
authors.forename: A
authors.initials: A
authors.lastname: Manoharan
authors.forename: P
authors.initials: P
authors.lastname: Narang
mesh_terms: D000151:Acinetobacter Infections / Q000382:microbiology; D040981:Acinetobacter baumannii / Q000187:drug effects / Q000201:enzymology*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D015378:Imipenem / Q000494:pharmacology; D000077731:Meropenem; D008826:Microbial Sensitivity Tests / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D013845:Thienamycins / Q000494:pharmacology; D018440:beta-Lactam Resistance; D001618:beta-Lactamases / Q000235:genetics* / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D013845:Thienamycins; D015378:Imipenem; D001618:beta-Lactamases; C063912:carbapenemase; D000077731:Meropenem