Journal of Antimicrobial Chemotherapy · 2014 · 37 citations · 14 references
Life SciencesGeneticsMicrobiologyAntimicrobial ResistanceBacteriologyMedical MicrobiologyAntibiotic ResistanceAntibiotic Resistance GeneticsBacterial GeneticsBeta-lactam ResistanceResistance Gene DiscoveryResistance GenesBeta-lactamase GenesCephalosporin ResistanceCloningConjugationExtended-spectrum Beta-lactamase PlasmidsExtended-spectrum Beta-lactamase-producing OrganismsExtended-spectrum Beta-lactamasesResistance In Clinical Isolates
OBJECTIVES: To characterize a novel CTX-M chimera, CTX-M-137, from Escherichia coli clinical isolates in China. METHODS: Isolates were collected from five hospitals between 22 February 2009 and 20 December 2011. Resistance genes were investigated by PCR. blaCTX-M-137 was cloned and purified for kinetic measurements. Conjugation experiments, S1-PFGE and Southern blotting were performed to study the plasmid harbouring blaCTX-M-137. The genetic environment of blaCTX-M-137 was determined by genomic cloning and sequencing. RESULTS: A total of 247 cephalosporin-resistant E. coli were identified. blaCTX-M group genes were the most prevalent extended-spectrum β-lactamase (ESBL) genes, with 71 isolates harbouring blaCTX-M-1 group genes and 137 isolates harbouring blaCTX-M-9 group genes. A novel chimera of CTX-M-14-like and CTX-M-15-like ESBLs, designated CTX-M-137, was identified from a 60-year-old man with a urinary tract infection. The N-terminus of CTX-M-137 matched CTX-M-14 and the C-terminus matched CTX-M-15. CTX-M-137 conferred resistance to ceftazidime, cefotaxime and aztreonam. Purified CTX-M-137 showed good hydrolytic activity against ceftazidime and cefotaxime, and was inhibited by clavulanic acid. The blaCTX-M-137 was carried on an ∼83 kb IncI1 plasmid. blaCTX-M-137 was carried on a complete transposition unit ISEcp1-blaCTX-M-137-Δorf477 inserted into yagA, which is part of the IncI1 plasmid backbone. CONCLUSIONS: We identified a novel CTX-M chimera, CTX-M-137, with a CTX-M-14-like N-terminus and a CTX-M-15-like C-terminus. Our findings suggest an ongoing diversification of CTX-M-type ESBLs through recombination events.
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Identification of plasmids by PCR-based replicon typing
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