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All the 41 clinical isolates previously phenotypically catalogued by the confirmatory method recommended by CLSI (disc diffusion assay) as ESBL positive were correctly detected by FC analysis following staining with DIBAC4(3). Hence, DiBAC4(3) was found to be a sensitive probe for assessing the impairment of cell membrane potential resulting from cephalosporin action. Positive FC results were obtained irrespective of ESBL type. Interestingly, despite the presence of multiple ��-lactamases genes, a fact earlier described as impairing the effectiveness of the ��-lactamase inhibitor CLA, the FC detection of ESBL-positive strains was highly successful. Nevertheless, false negative results might Lapatinib clinical trial be obtained whenever rare ESBL types are present, because they are not inhibited by CLA [16]. False negative results could be also S6 Kinase expected with isolates expressing enzymes (Complex Mutant TEM), which confer resistance to cephalosporins but are partially inhibited by CLA [17]. In order to assess the true specificity of the method hereby reported, isolates with beta-lactamases other than ESBLs, such as VIM (class B carbapenemase), OXA (class D penicillinases) and DHA-1 (class C AmpC beta-lactamases), which are not inhibited by CLA (CLA index LY294002 research buy bacilli using FC, none of them reported the effect of its association with CLA in order to detect the presence of ESBLs [19�C25]. The estimated costs for consumables and reagents used for this new method for ESBL detection is rather low, around 2.5�� per sample (per microorganism) screened without taking into account the costs of the FC equipment and technician. However, such apparatus is frequently found in most hematology/immunology laboratories and could certainly be shared by microbiologists.