RESUMO
OBJECTIVES: The aim of this study was to characterize carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates recovered from clinical specimens at a tertiary care hospital in Egypt over a period of 15 months. METHODS: Eight CRKP isolates were included in this study. The minimum inhibitory concentrations of different antibiotics were determined by broth microdilution and Etest methods. Multilocus sequence typing was performed. Antibiotic resistance genes were assessed by PCR and DNA sequencing. Plasmid analysis was done by S1 nuclease digestion of whole genomic DNA followed by pulsed-field gel electrophoresis (S1-PFGE). RESULT: Eight carbapenem-resistant NDM-1-producing K. pneumoniae isolates of three different sequence types (ST) were identified (ST147, ST11, and ST17), in which blaNDM-1 was carried by either IncR or untypeable plasmids. Seven out of the eight isolates also contained the rmtF methylase gene. CONCLUSION: This study describes the occurrence of IncR plasmids carrying blaNDM-1 and rmtF in Egypt, raising concerns regarding this type of replicon and its role in the transmission of these resistance determinants.
Assuntos
Antibacterianos/farmacologia , Carbapenêmicos/farmacologia , Resistência Microbiana a Medicamentos/efeitos dos fármacos , Infecções por Enterobacteriaceae/tratamento farmacológico , Infecções por Klebsiella/tratamento farmacológico , Klebsiella pneumoniae/efeitos dos fármacos , beta-Lactamases/metabolismo , Resistência Microbiana a Medicamentos/genética , Egito/epidemiologia , Eletroforese em Gel de Campo Pulsado , Enterobacteriaceae/isolamento & purificação , Infecções por Enterobacteriaceae/epidemiologia , Infecções por Enterobacteriaceae/microbiologia , Humanos , Infecções por Klebsiella/epidemiologia , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/genética , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Tipagem de Sequências Multilocus , Plasmídeos , Análise de Sequência de DNA , beta-Lactamases/efeitos dos fármacos , beta-Lactamases/genéticaRESUMO
An investigation was carried out into the genetic mechanisms responsible for multidrug resistance in nine carbapenem-resistant Pseudomonas aeruginosa isolates from different hospitals in Recife, Brazil. Susceptibility to antimicrobial agents was determined by broth microdilution. Polymerase chain reaction (PCR) was employed to detect the presence of genes encoding ß-lactamases, aminoglycoside-modifying enzymes (AMEs), 16S rRNA methylases, integron-related genes and OprD. Expression of genes coding for efflux pumps and AmpC cephalosporinase were assessed by quantitative PCR. The outer membrane proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The blaSPM-1, blaKPC-2 and blaGES-1 genes were detected in P. aeruginosa isolates in addition to different AME genes. The loss of OprD in nine isolates was mainly due to frameshift mutations, premature stop codons and point mutations. An association of loss of OprD with the overexpression of MexAB-OprM and MexXY-OprM was observed in most isolates. Hyper-production of AmpC was also observed in three isolates. Clonal relationship of the isolates was determined by repetitive element palindromic-PCR and multilocus sequence typing. Our results show that the loss of OprD along with overexpression of efflux pumps and ß-lactamase production were responsible for the multidrug resistance in the isolates analysed.
Assuntos
Carbapenêmicos/metabolismo , Farmacorresistência Bacteriana Múltipla/genética , Mutação , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/genética , Resistência beta-Lactâmica/genética , beta-Lactamases/metabolismo , Aminoglicosídeos/metabolismo , Anfotericina B/análogos & derivados , Anfotericina B/metabolismo , Antifúngicos/metabolismo , Brasil , Cefalosporinase/classificação , Cefalosporinase/metabolismo , Códon sem Sentido/metabolismo , Ativação Enzimática/genética , Mutação da Fase de Leitura/genética , Regulação Bacteriana da Expressão Gênica/genética , Humanos , Proteínas de Membrana Transportadoras/metabolismo , Metiltransferases/metabolismo , Nucleotidiltransferases/metabolismo , Mutação Puntual/genética , Porinas/metabolismo , Pseudomonas aeruginosa/enzimologia , Pseudomonas aeruginosa/isolamento & purificação , Sequências Repetitivas de Ácido Nucleico , beta-Lactamases/genéticaRESUMO
An investigation was carried out into the genetic mechanisms responsible for multidrug resistance in nine carbapenem-resistant Pseudomonas aeruginosaisolates from different hospitals in Recife, Brazil. Susceptibility to antimicrobial agents was determined by broth microdilution. Polymerase chain reaction (PCR) was employed to detect the presence of genes encoding β-lactamases, aminoglycoside-modifying enzymes (AMEs), 16S rRNA methylases, integron-related genes and OprD. Expression of genes coding for efflux pumps and AmpC cephalosporinase were assessed by quantitative PCR. The outer membrane proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The blaSPM-1, blaKPC-2 and blaGES-1 genes were detected in P. aeruginosaisolates in addition to different AME genes. The loss of OprD in nine isolates was mainly due to frameshift mutations, premature stop codons and point mutations. An association of loss of OprD with the overexpression of MexAB-OprM and MexXY-OprM was observed in most isolates. Hyper-production of AmpC was also observed in three isolates. Clonal relationship of the isolates was determined by repetitive element palindromic-PCR and multilocus sequence typing. Our results show that the loss of OprD along with overexpression of efflux pumps and β-lactamase production were responsible for the multidrug resistance in the isolates analysed.
