title: Use of extracts versus whole-cell bacterial suspensions in the identification of Staphylococcus aureus beta-lactamase variants.
abstract: We previously have shown that extracts of S. aureus isolates which produce the recognized serotypes of staphylococcal beta-lactamase (A, B, C, D) differ in the rates at which they hydrolyze selected cephalosporins, exhibiting substrate profiles which are distinctive for each serotype. In an effort to simplify the methods employed in identifying the different staphylococcal beta-lactamases, we evaluated whether distinctive substrate profiles could be obtained by using whole-cell suspensions of 115 beta-lactamase-producing isolates of S. aureus. Compared with extracts from the same strains, the whole-cell bacterial suspensions not only were simpler to prepare but enabled beta-lactamase typing of a higher proportion of the evaluated strains (86 versus 97%, respectively). Furthermore, the use of whole-cell bacterial suspensions enabled the simultaneous quantitation of the beta-lactamase activity exhibited by each strain. Additionally, by comparing the quantitative activity of beta-lactamase-induced and -uninduced preparations of the same strain, induction ratios (i.e., induced/uninduced activity) could be derived, yielding information regarding the regulation of beta-lactamase production by each strain. We believe that the utilization of whole-cell methods, such as those employed in this study, will facilitate the investigation of qualitative and quantitative differences in beta-lactamase production among clinical and reference isolates of S. aureus.
authors.affiliation: Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee 37232.
authors.forename: D S
authors.initials: DS
authors.lastname: Kernodle
authors.forename: P A
authors.initials: PA
authors.lastname: McGraw
authors.forename: C W
authors.initials: CW
authors.lastname: Stratton
authors.forename: A B
authors.initials: AB
authors.lastname: Kaiser
mesh_terms: D002474:Cell-Free System; D002478:Cells, Cultured; D002511:Cephalosporins / Q000378:metabolism; D014644:Genetic Variation; D006868:Hydrolysis; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D008722:Methods; D013211:Staphylococcus aureus / Q000201:enzymology / Q000235:genetics; D013379:Substrate Specificity; D013535:Suspensions; D001618:beta-Lactamases / Q000235:genetics / Q000378:metabolism*
chemical_list: D002511:Cephalosporins; D007527:Isoenzymes; D013535:Suspensions; D001618:beta-Lactamases; C021720:nitrocefin