First Report of an OXA-48- and CTX-M-213-Producing Kluyvera Species Clone Recovered from Patients Admitted in a University Hospital in Madrid, Spain

Abstract

species other than also contribute to OXA-48 carbapenemase endemicity. We studied the emergence of an OXA-48-producing species clone, which expresses the novel CTX-M-213 enzyme, colonizing patients in our hospital. Rectal swabs from patients admitted in four wards (March 2014 to March 2016; R-GNOSIS project) were seeded onto Chromo ID-ESBL) and Chrom-CARB/OXA-48 chromogenic agar plates. Carbapenemases and extended-spectrum β-lactamases (ESBLs) were characterized (PCR, sequencing, cloning, and site-directed mutagenesis), and antibiotic susceptibility was determined. Clonal relatedness was established (XbaI pulsed-field gel electrophoresis [XbaI-PFGE]), and plasmid content was studied (transformation, S1 nuclease digestion-PFGE, SB-hybridization, restriction fragment length polymorphism [RFLP] analysis [DraI and HpaI], and PCR [incompatibility group and A,U, and A genes]). Whole-genome sequencing (WGS) (Illumina HiSeq-2500) and further bioinformatics analysis of plasmids (PLACNET and plasmidSPAdes) were performed. Patients' charts were reviewed. Six unrelated patients (median age, 75 years [range, 59 to 81 years]; 4/6 male patients) colonized with OXA-48-producing species isolates (>95% similarity of the PFGE pattern) were identified. Nosocomial acquisition was demonstrated. In two patients, OXA-48-producing species isolates coexisted with OXA-48-producing, , and The gene was located on an ∼60-kb IncL plasmid related to IncL/M-pOXA-48a and the novel gene in a conserved chromosomal region of species isolates. CTX-M-213, different from CTX-M-13 (K56E) but conferring a similar β-lactam resistance profile, was identified. Genomic analysis also revealed a 177-kb IncF plasmid (class I integron harboring and) and an 8-kb IncQ plasmid (IS- ). We describe the first plasmid in spp. and the novel chromosomal CTX-M-213 enzyme and highlight further nosocomial dissemination of through clonal lineages or plasmids related to IncL/M-pOXA-48a.

The Global Ascendency of OXA-48-Type Carbapenemases

“…Subsequently OXA-48 was reported from several members of the Enterobacterales, especially E. coli (108), the E. cloacae complex (109), Citrobacter freundii (98), Serratia marcescens (74), Proteus mirabilis (110), Kluyvera spp. (111), Klebsiella oxytoca (112), and Salmonella spp. (113).…”

<p><em>Raoultella ornithinolytica</em>: Emergence and Resistance</p>

“…50,51 Other Raoultella species were also described as exhibiting OXA-48 carbapenemase. [52][53][54][55] Nonetheless, a few strains of R. ornithinolytica generating simultaneously different types of beta-lactamase (TEM, SHV, KPC and OXA) were described. 42,56 The first case of co-existence of bla KPC-2 and bla IMP-4 carbapenemase genes in the genus Raoultella was reported by Zheng et al (2015) in a 13-year-old male patient with postoperative infection after an orthopedic surgery.…”

The first case of third-generation cephalosporins resistant Kluyvera ascorbata biliary tract infection in Japan: A case report and review of the literature

“…Recently, CTX-M type β-lactamase is described as the most prevalent variant of ESBLs [12]. Moreover, the mobilization of chromosomal bla CTX-M gene is occasionally reported; plasmid encoding CTX-M genes could cause emerging multi-drug resistant pathogen [15].…”