Rapid method of extraction and analysis of extended-spectrum beta-lactamases from clinical strains of Klebsiella pneumoniae.
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.
Paterson
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Louis Stokes Veterans Affairs Medical Center, 10701 East Blvd, Cleveland, OH 44106, USA.
Rice
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Bonomo
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D006801:Humans; D007525:Isoelectric Focusing; D007711:Klebsiella pneumoniae / Q000201:enzymology*; D009113:Muramidase; D001618:beta-Lactamases / Q000032:analysis / Q000302:isolation & purification*
D009113:Muramidase; D001618:beta-Lactamases