Open AccessEvaluation of a New Etest for Detecting Metallo-β-Lactamases in Routine Clinical Testing
Author(s) -
Timothy R. Walsh,
Anne Bolmström,
Anette Qwärnström,
Ana Cristina Gales
Publication year - 2002
Publication title -
journal of clinical microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.349
H-Index - 255
eISSN - 1070-633X
pISSN - 0095-1137
DOI - 10.1128/jcm.40.8.2755-2759.2002
Subject(s) - etest , microbiology and biotechnology , agar , proteus mirabilis , agar dilution , biology , bacteroides fragilis , imipenem , bacteria , minimum inhibitory concentration , pseudomonas aeruginosa , antimicrobial , antibiotics , antibiotic resistance , genetics
Several Etest (AB BIODISK, Solna, Sweden) gradient formats were developed for detection of metallo-beta-lactamases based on the reduction of imipenem (IP) or ceftazidime (TZ) MICs in the presence of EDTA or 2-mercaptopropionic acid (MPA). The Etest metallo-beta-lactamase (Etest MBL) strips consisted of a double-sided seven-dilution range of IP or TZ (4 to 256 microg/ml) and IP or TZ (1 to 64 microg/ml) overlaid with a constant concentration of EDTA or MPA. The prototype strips were evaluated on several agar media (brain heart infusion agar, Isosensitest agar, nutrient agar, and Mueller-Hinton agar for aerobes and brucella blood agar for anaerobes) with 138 challenge strains: Acinetobacter spp. (n = 9), Aeromonas spp. (n = 8), Chryseobacterium spp. (n = 28), Escherichia coli (n = 1), Klebsiella pneumoniae (n = 4), Pseudomonas aeruginosa (n = 14), Proteus mirabilis (n = 3), Serratia spp. (n = 10), Stenotrophomonas maltophilia (n = 43), Sphingobacterium spp. (n = 3), and Bacteroides fragilis group (n = 15). PCR analysis using specific primers for IMP-1, L1, CcrA, and bla(B/C) confirmed the presence of the metallo-beta-lactamase genes. Enzyme assays were also performed with IP as an indicator substrate followed by EDTA inhibition profiles. EDTA was found to be a better inhibitor of metallo-beta-lactamases, especially for anaerobes. IP was a better than TZ. Mueller-Hinton agar was the preferred medium, particularly when compared to Isosensitest agar, which frequently produced falsely low MICs for IP. Etest IP plus IP-EDTA with Mueller-Hinton agar had a sensitivity of 94% (79 of 84) and specificity of 95% (124 of 130). The Etest MBL strip appears to be an acceptable diagnostic reagent to detect metallo-beta-lactamase phenotypes in the clinical microbiology laboratory.