Evaluation of Different Methods for Detection of Metallo-beta-Lactamases in Pseudomonas aeruginosa Clinical Isolates

Abstract

Metallo-beta-lactamases (MBLs) produced by Pseudomnonas aeruginosa are a serious threat due to their ability to be transmitted between the same as well as different bacterial species. Different methods are applied in the clinical laboratory to detect MBLs. The aim of this study was to compare 4 phenotypic methods and a PCR assay for their ability to detect MBLs in clinical isolates of carbapenem-resistant P. aeruginosa strains. The study embraced a total of 70 carbapenem-resistant P. aeruginosa strains isolated in The Department of Microbiology of Dr. A. Jurasz University Hospital in Bydgoszcz. The highest percentage (42.9%) of the strains were isolated from Intensive Care Unit patients, mainly from urine samples (31.4%). Methods used in this study were: double-disc synergy tests in two combinations: using ceftazidime with 2-mercaptopropionic acid and imipenem with EDTA, differences in inhibition zone diameters between discs with imipenem/EDTA and imipenem, Etest MBL (AB Biodisk) and molecular amplification of bla(IMP) and bla(VIM) genes responsible for producing MBLs, using PCR assay. The lowest percentage (1.4%) of positive results in detection of MBLs was obtained using PCR assay, the highest (72.9%) by double-disc synergy tests with imipenem and EDTA, but the specificity of this method may be low.