ABSTRACT
Abstract Antimicrobial resistance due to carbapenemase production in Enterobacteriaceaeclinical isolates is a global threat. Klebsiella pneumoniae harboring the blaKPCgene is one ofthe major concerns in hospital settings in Latin America.The aim of this study was to characterize the antibiotic resistance mechanisms and to typifyfour carbapenem-resistant K. pneumoniae clinical isolates from the city of Manizales, Colombia.We identified blaKPC-3in all four isolates by polymerase chain reaction and subsequentsequencing. The plasmid-mediated quinolone resistance genes qnrB19-like and aac(6)Ib-cr;fosfomycin resistance gene fosA and an insertion sequence IS5-like in mgrB (colistin resistance)were also detected. Sequence types ST11 with capsular type wzi75, and ST258 with wzi154,were characterized. The blaKPC-3gene was mobilized in a 100-kb IncFIB conjugative plasmidwith vagCD toxin-antitoxin system.This work reports multiple resistance genes in blaKPC-producing K. pneumoniae and the firstoccurrence of ST11 clinical isolates harboring blaKPC-3in Latin America.
Resumen La resistencia a antibióticos mediada por la producción de carbapenemasas en aislamientos clínicos de Enterobacteriaceae es una amenaza mundial. Klebsiella pneumoniae portador de blaKPC es uno de los mayores problemas a nivel hospitalario en Latinoamérica. El objetivo de este estudio fue caracterizar los mecanismos de resistencia antibiótica y tipificar cuatro aislamientos clínicos de K. pneumoniae resistentes a carbapenems obtenidos en la ciudad de Manizales, Colombia. Se identificó blaKPC-3 en todos los aislamientos mediante reacción en cadena de polimerasa y secuenciación. También se detectaron los genes de resistencia transferible a quinolonas qnrB19-like y aac(6')Ib-cr y a fosfomicina fosA, y la secuencia de inserción /S5-like en mgrB (asociada a la resistencia a colistina). Se caracterizaron los secuenciotipos ST11 (cápsula wzi75) y ST258 (cápsula wzi154). Se comprobó que blaKPC-3 fue movilizado por un plásmido conjugativo IncFIB-vagCD de 100kb. En este trabajo se reportan múltiples genes de resistencia en K. pneumoniae productor de blaKPC y se describen por primera vez aislamientos clínicos ST11 productores de blaKPC-3 en Latinoamérica.
Subject(s)
Humans , Klebsiella Infections/microbiology , Klebsiella pneumoniae/isolation & purification , Bacterial Proteins/genetics , beta-Lactamases/genetics , Microbial Sensitivity Tests , Klebsiella pneumoniae/genetics , Latin America/epidemiology , Anti-Bacterial Agents/pharmacologyABSTRACT
Antimicrobial resistance due to carbapenemase production in Enterobacteriaceae clinical isolates is a global threat. Klebsiellapneumoniae harboring the blaKPC gene is one of the major concerns in hospital settings in Latin America. The aim of this study was to characterize the antibiotic resistance mechanisms and to typify four carbapenem-resistant K. pneumoniae clinical isolates from the city of Manizales, Colombia. We identified blaKPC-3 in all four isolates by polymerase chain reaction and subsequent sequencing. The plasmid-mediated quinolone resistance genes qnrB19-like and aac(6')Ib-cr; fosfomycin resistance gene fosA and an insertion sequence IS5-like in mgrB (colistin resistance) were also detected. Sequence types ST11 with capsular type wzi75, and ST258 with wzi154, were characterized. The blaKPC-3 gene was mobilized in a 100-kb IncFIB conjugative plasmid with vagCD toxin-antitoxin system. This work reports multiple resistance genes in blaKPC-producing K. pneumoniae and the first occurrence of ST11 clinical isolates harboring blaKPC-3 in Latin America.
Subject(s)
Klebsiella Infections , Klebsiella pneumoniae/isolation & purification , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Humans , Klebsiella Infections/microbiology , Klebsiella pneumoniae/genetics , Latin America/epidemiology , Microbial Sensitivity Tests , beta-Lactamases/geneticsABSTRACT
OBJECTIVES: Since KPC-2-producing Klebsiella pneumoniae are associated with successful dissemination of a major clone, defined as sequence type 258 (ST258), the aim of this study was to perform whole-genome sequencing (WGS) of the first colistin-resistant K. pneumoniae strain (Kpn666) carrying blaKPC-2 identified in Uruguay in 2011 in order to identify genomic and phylogenetic traits. METHODS: WGS of strain Kpn666 isolated from an asymptomatic urinary tract infection was performed using Illumina MiSeq, and de novo assembly was performed using SPADES v.3.11. Contigs were re-ordered using the ST258 reference genome NJST258_1 (GenBank CP006923) and were oriented with the MAUVE Contig Mover. Twenty complete genomes of K. pneumoniae identified as ST258 using the Pasteur MLST site were downloaded from GenBank (May 2017). A maximum-likelihood tree was created using MEGA7 based on core single nucleotide polymorphisms (SNPs) from whole-genome alignment obtained with SNP sites (https://github.com/sanger-pathogens/snp-sites). RESULTS: WGS analysis revealed a genome of 5448179bp (5232 CDS, 108 RNAs). Phylogenetic analysis identified that Kpn666 belonged to clade I lineage of ST258. Further studies also identified IncR, IncFIB(K) and IncFII(K) plasmid replicons and 11 transferable associated antimicrobial resistance genes (ARGs) comprising four drug classes. The mgrB gene involved in colistin resistance was shown to be disrupted by insertion of an IS5-like element. CONCLUSIONS: The first isolate of KPC-2-producing K. pneumoniae detected in Uruguay was sequenced and the results confirm the ability of this bacterium to capture several ARGs. The KPC-2 carbapenemase in Uruguay is likely to have been introduced by the high-risk clone ST258.
