RESUMO
The increasing prevalence of third-generation cephalosporin-resistant Enterobacteriaceae is a worldwide problem. Recent studies showed that poultry meat and humans share identical Extended-Spectrum Beta-Lactamase genes, plasmid types, and Escherichia coli strain types, suggesting that transmission from poultry meat to humans may occur. The aim of this study was to compare plasmid-encoded Ambler class C beta-lactamase (pAmpC) genes, their plasmids, and bacterial strain types between E. coli isolates from retail chicken meat and clinical isolates in the Netherlands. In total, 98 Dutch retail chicken meat samples and 479 third-generation cephalosporin non-susceptible human clinical E. coli isolates from the same period were screened for pAmpC production. Plasmid typing was performed using PCR-based replicon typing (PBRT). E coli strains were compared using Multi-Locus-Sequence-Typing (MLST). In 12 of 98 chicken meat samples (12%), pAmpC producing E. coli were detected (all blaCMY-2). Of the 479 human E. coli, 25 (5.2%) harboured pAmpC genes (blaCMY-2 n = 22, blaACT n = 2, blaMIR n = 1). PBRT showed that 91% of poultry meat isolates harboured blaCMY-2 on an IncK plasmid, and 9% on an IncI1 plasmid. Of the human blaCMY-2 producing isolates, 42% also harboured blaCMY-2 on an IncK plasmid, and 47% on an IncI1 plasmid. Thus, 68% of human pAmpC producing E. coli have the same AmpC gene (blaCMY-2) and plasmid type (IncI1 or IncK) as found in poultry meat. MLST showed one cluster containing one human isolate and three meat isolates, with an IncK plasmid. These findings imply that a foodborne transmission route of blaCMY-2 harbouring plasmids cannot be excluded and that further evaluation is required.
Assuntos
Proteínas de Bactérias/genética , Infecções por Escherichia coli/microbiologia , Escherichia coli/genética , Carne/microbiologia , Plasmídeos/genética , beta-Lactamases/genética , Animais , Antibacterianos/farmacologia , Escherichia coli/classificação , Escherichia coli/efeitos dos fármacos , Humanos , Tipagem de Sequências Multilocus , Países Baixos , Filogenia , Reação em Cadeia da Polimerase , Aves DomésticasRESUMO
Extended-spectrum ß-lactamases (ESBLs) are emerging worldwide, making rapid and adequate ESBL detection crucial for infection control measures as well as for the choice of correct antimicrobial therapy. The aim of this study was to compare the performance of a novel rapid ligation-mediated real-time PCR (LM-PCR) with a combination disc test (CDT). In total, 172 prospective putative ESBL-positive Enterobacteriaceae isolates from clinical specimens based on VITEK2 results were included in this study and tested with the phenotypic CDT and the LM-PCR. Positive ESBL results were obtained in 100 and 95 isolates using CDT and LM-PCR, respectively. The sensitivity, specificity, negative predictive value and positive predictive value of the LM-PCR were 99.0, 92.2, 98.6 and 94.0â%, respectively, compared with the CDT. The LM-PCR technique provides an important reduction in turnaround time (~4.5 h versus overnight incubation using CDT) for ESBL confirmation. As a consequence, all ESBL results are available within the same day, making this assay an important tool for rapid and accurate ESBL detection.
Assuntos
Enterobacteriaceae/enzimologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , beta-Lactamases/genética , beta-Lactamases/metabolismo , Antibacterianos/farmacologia , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Farmacorresistência Bacteriana Múltipla/genética , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/genética , Variação Genética , Fenótipo , Valor Preditivo dos Testes , Reação em Cadeia da Polimerase em Tempo Real/economia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Fatores de TempoRESUMO
There is a global increase in infections caused by Enterobacteriaceae with plasmid-borne ß-lactamases that confer resistance to third-generation cephalosporins. The epidemiology of these bacteria is not well understood, and was, therefore, investigated in a selection of 636 clinical Enterobacteriaceae with a minimal inhibitory concentration >1 mg/L for ceftazidime/ceftriaxone from a national survey (75% E. coli, 11% E. cloacae, 11% K. pneumoniae, 2% K. oxytoca, 2% P. mirabilis). Isolates were investigated for extended-spectrum ß-lactamases (ESBLs) and ampC genes using microarray, PCR, gene sequencing and molecular straintyping (Diversilab and multi-locus sequence typing (MLST)). ESBL genes were demonstrated in 512 isolates (81%); of which 446 (87%) belonged to the CTX-M family. Among 314 randomly selected and sequenced isolates, bla(CTX-M-15) was most prevalent (nâ=â124, 39%), followed by bla(CTX-M-1) (nâ=â47, 15%), bla(CTX-M-14) (nâ=â15, 5%), bla(SHV-12) (nâ=â24, 8%) and bla(TEM-52) (nâ=â13, 4%). Among 181 isolates with MIC ≥16 mg/L for cefoxitin plasmid encoded AmpCs were detected in 32 and 27 were of the CMY-2 group. Among 102 E. coli isolates with MIC ≥16 mg/L for cefoxitin ampC promoter mutations were identified in 29 (28%). Based on Diversilab genotyping of 608 isolates (similarity cut-off >98%) discriminatory indices of bacteria with ESBL and/or ampC genes were 0.994, 0.985 and 0.994 for E. coli, K. pneumoniae and E. cloacae, respectively. Based on similarity cut-off >95% two large clusters of E. coli were apparent (of 43 and 30 isolates) and 21 of 21 that were typed by belonged to ST131 of which 13 contained bla(CTX-M-15). Our findings demonstrate that bla(CTX-M-15) is the most prevalent ESBL and we report a larger than previously reported prevalence of ampC genes among Enterobacteriaceae responsible for resistance to third-generation cephalosporins.
Assuntos
Antibacterianos/farmacologia , Resistência às Cefalosporinas/genética , Cefalosporinas/farmacologia , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/genética , beta-Lactamases/genética , Adolescente , Adulto , Idoso , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana , Criança , Pré-Escolar , Enterobacteriaceae/classificação , Humanos , Lactente , Recém-Nascido , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Países Baixos , Adulto JovemRESUMO
In 271 Enterobacter blood culture isolates from 12 hospitals, extended-spectrum beta-lactamase (ESBL) prevalence varied between 0% and 30% per hospital. High prevalence was associated with dissemination, indicating the potential relevance of infection control measures. Screening with cefepime or Vitek 2, followed by a cefepime/cefepime-clavulanate Etest, was an accurate strategy for ESBL detection in Enterobacter isolates (positive predictive value, 100%; negative predictive value, 99%).
Assuntos
Antibacterianos/farmacologia , Técnicas de Laboratório Clínico/métodos , Enterobacter/enzimologia , beta-Lactamases/biossíntese , beta-Lactamas/farmacologia , Bacteriemia/microbiologia , Enterobacter/isolamento & purificação , Infecções por Enterobacteriaceae/microbiologia , Humanos , Testes de Sensibilidade Microbiana/métodos , Valor Preditivo dos TestesAssuntos
Infecções por Escherichia coli/microbiologia , Escherichia coli/enzimologia , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/enzimologia , beta-Lactamases/metabolismo , Adulto , Idoso , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Carbapenêmicos/farmacologia , Carbapenêmicos/uso terapêutico , DNA Bacteriano/química , DNA Bacteriano/genética , Escherichia coli/genética , Infecções por Escherichia coli/tratamento farmacológico , Feminino , Humanos , Masculino , Testes de Sensibilidade Microbiana , Países Baixos , Reação em Cadeia da Polimerase , Infecções Estafilocócicas/tratamento farmacológico , Staphylococcus aureus/efeitos dos fármacos , ViagemRESUMO
This is the first report of 3 patients in whom carbapenemase-producing Klebsiella pneumoniae was identified in the Netherlands following foreign travel. They were a 55-year-old man who had undergone chemotherapy for lung cancer metastases, a 66-year-old woman and a 30-year-old man. The first patient was transferred from a Greek hospital; his isolate belonged to an epidemic clone (multilocus sequence type 258) with a KPC-2 carbapenemase gene. The patient died from pneumonia. The other two patients, who had been travelling around in India, were found to be colonised in the gasto-intestinal tract with different multiresistant K. pneumoniae isolates containing a New Delhi metallo-carbapenemase gene (NDM-1). The rapid emergence and dissemination of Enterobacteriaceae resistant to carbapenems such as imipenem and meropenem poses a considerable threat to clinical patient care and public health. Carbapenemase-producing strains are characterized by resistance to nearly all available beta-lactam antibiotics including cephalosporins and carbapenems. These strains are often also resistant to other classes of antibiotics. Invasive infections by these strains are associated with high morbidity and mortality rates. Adequate microbiological laboratory detection and infection control measures in hospital are pivotal to preventing dissemination in the Dutch healthcare setting.