Sensitive EDTA-based microbiological assays for detection of metallo-β-lactamases in nonfermentative gram-negative bacteria

被引:55
作者
Marchiaro, P
Mussi, MA
Ballerini, V
Pasteran, F
Viale, AM
Vila, AJ
Limansky, AS
机构
[1] Univ Nacl Rosario, Inst Biol Mol & Celular Rosario, Dept Microbiol, Fac Ciencias Bioquim & Farmaceut, RA-2000 Rosario, Santa Fe, Argentina
[2] Univ Nacl Rosario, Inst Biol Mol & Celular Rosario, Dept Quim Biol, Fac Ciencias Bioquim & Farmaceut, RA-2000 Rosario, Santa Fe, Argentina
[3] Ctr Especialidades Med Ambulatorias Rosario, Dept Bioquim Municipal, Rosario, Argentina
[4] ANLIS, Serv Antimicrobianos, Dept Bacteriol, Ciudad Auton Buenos Aires, Buenos Aires, DF, Argentina
关键词
D O I
10.1128/JCM.43.11.5648-5652.2005
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The worldwide spread of metallo-beta-lactamase (MBL)-producing gram-negative bacilli represents a great concern nowadays. Sensitive assays for their specific detection are increasingly demanded to aid infection control and to prevent their dissemination. We have developed a novel microbiological assay employing crude bacterial extracts, designated EDTA-imipenem microbiological assay (EIM), to identify MBLs in nonfermentative gram-negative clinical strains. We also evaluated the ability of EIM to detect MBLs in comparison to those of other currently employed screening methods, such as the EDTA disk synergy test (EDS) with imipenem as a substrate and the Etest method. The sensitivities of EIM and Etest were similar (1 versus 0.92, respectively) and much higher than that of EDS (0.67). Moreover, both EIM and Etest displayed the maximum specificity. Modifications were introduced to EDS, including the simultaneous testing of three different beta-lactams (imipenem, meropenem, and ceftazidime) and two different EDTA concentrations. This resulted in a sensitivity improvement (0.92), albeit at a cost to its specificity. A simple strategy to accurately detect MBL producers is proposed; this strategy combines (i) an initial screening of the isolates by the extended EDS assay to select the potential candidates and (ii) confirmation of the true presence of MBL activity by EIM.
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页码:5648 / 5652
页数:5
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