JAC Advance Access originally published online on May 5, 2004
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Journal of Antimicrobial Chemotherapy (2004) 53, 1076-1080
© 2004 The British Society for Antimicrobial Chemotherapy
Citrobacter koseri and Citrobacter amalonaticus isolates carry highly divergent ß-lactamase genes despite having high levels of biochemical similarity and 16S rRNA sequence homology
Bristol Centre for Antimicrobial Research and Evaluation, Department of Pathology & Microbiology, University of Bristol, School of Medical Sciences, University Walk, Bristol BS8 1TD, UK
Received 25 February 2004; accepted 12 March 2004
Objectives: Isolates previously identified as Citrobacter diversus are now known as Citrobacter koseri. We measured sequence variation at the ß-lactamase structural gene among a group of clinical isolates originally identified as C. diversus by API 20E profiling.
Methods: ß-Lactamase and 16S rRNA genes were amplified by PCR and sequenced by standard methods. ß-Lactamase induction was attempted in liquid-grown cultures using cefoxitin. Nitrocefin hydrolysis assays were performed using a spectrophotometer.
Results: Analysis of 16S rRNA gene sequences showed that Citrobacter spp. isolates with an inducible ß-lactamase gene, cdiA, closely related to C. koseri NF85 and ULA27 are actually Citrobacter amalonaticus. C. koseri isolates, whose identities were confirmed by 16S rRNA sequencing, produce a class A ß-lactamase, Cko, constitutively at low levels. The cko and cdiA ß-lactamase genes share <45% identity.
Conclusions: We have confirmed that cko is a ß-lactamase gene carried by C. koseri, and that isolates previously identified as C. koseri , but carrying the cdiA ß-lactamase gene are C. amalonaticus. Thus, ß-lactamase-gene-specific PCR may provide a valuable tool to differentiate these biochemically homogeneous Citrobacter species.
Keywords: Citrobacter, ß-lactamases, phylogenetics, induction
* Corresponding author. Tel: +44-117-9287897; Fax: +44-117-9287896; E-mail: Matthewb.Avison{at}bris.ac.uk
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