title: Rapid method of extraction and analysis of extended-spectrum beta-lactamases from clinical strains of Klebsiella pneumoniae.
abstract: The extraction of periplasmic beta-lactamases from Gram-negative bacilli is a necessary preliminary step to analytical isoelectric focusing. Previously described methods are time-consuming and require large amounts of broth. We describe a lysozyme-based method which needs just 5 mL broth and requires less than 24 h to perform. The method was reproducible in extracting beta-lactamases from reference strains containing known beta-lactamases. We applied the method to a collection of more than 70 extended-spectrum beta-lactamase-producing isolates from a multinational study of bacteremic isolates of Klebsiella pneumoniae. Further studies are being undertaken to assess the method's applicability to other bacterial species.
authors.affiliation: Louis Stokes Veterans Affairs Medical Center, 10701 East Blvd, Cleveland, OH 44106, USA.
authors.forename: D L
authors.initials: DL
authors.lastname: Paterson
authors.forename: L B
authors.initials: LB
authors.lastname: Rice
authors.forename: R A
authors.initials: RA
authors.lastname: Bonomo
mesh_terms: D006801:Humans; D007525:Isoelectric Focusing; D007711:Klebsiella pneumoniae / Q000201:enzymology*; D009113:Muramidase; D001618:beta-Lactamases / Q000032:analysis / Q000302:isolation & purification*
chemical_list: D009113:Muramidase; D001618:beta-Lactamases