Evaluation of different laboratory tests for the detection of metallo-beta-lactamase production in Enterobacteriaceae.
OBJECTIVE
Clinical isolates of Klebsiella pneumoniae (91), Escherichia coli (49), Enterobacter spp. (27), Proteus mirabilis (17), Citrobacter freundii (2), Providencia stuartii (3) and Serratia spp. (5), with various MICs of imipenem, were examined for production of metallo-beta-lactamases (MBLs) with different phenotypic laboratory tests that have been previously published to detect MBLs in Pseudomonas aeruginosa and Acinetobacter spp.


METHODS
A total of 194 (95 MBL-positive and 99 MBL-negative) clinical isolates with imipenem MICs < or = 0.25 to > 256 mg/L were examined. All isolates were evaluated by the double-disc synergy test (DDST), the combination disc test (CDT), the MBL Etest and the modified Hodge test. The presence of bla(VIM) and bla(IMP) genes was evaluated by in situ hybridization with specific probes and was certified by PCR.


RESULTS
In 30 bla(VIM)-positive isolates that exhibited MICs of imipenem < or = 4 mg/L, MBL Etest could not be evaluated. CDT with ceftazidime and 1900 or 750 microg of EDTA, and DDST after applying an imipenem disc 10 mm apart from a disc containing approximately 1900 microg of EDTA, showed the highest sensitivity (97.9% to 100%) and specificity (87.9% to 96%) rates among the analysed procedures. CDT with imipenem and 1900 microg of EDTA exhibited a sensitivity of 94.7% and showed very good specificity (98%).


CONCLUSIONS
The CDT with imipenem/imipenem+0.5 M EDTA or ceftazidime/ceftazidime+0.2 M EDTA and the DDST with imipenem 10 mm apart from EDTA are the most effective methods for the detection of MBLs in Enterobacteriaceae.
Galani
Irene
I
4th Department of Internal Medicine, Molecular Biology Section, University of Athens Medical School, Athens, Greece. egalani@med.uoa.gr
Rekatsina
Panagiota Danai
PD
Hatzaki
Despina
D
Plachouras
Diamantis
D
Souli
Maria
M
Giamarellou
Helen
H
D019411:Clinical Laboratory Techniques / Q000592:standards; D004755:Enterobacteriaceae / Q000201:enzymology* / Q000302:isolation & purification; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods / Q000592:standards*; D001618:beta-Lactamases / Q000096:biosynthesis* / Q000302:isolation & purification
D001618:beta-Lactamases