RESUMO
Enterobacteria-producing extended-spectrum ß-lactamases (ESBL) play an important role in healthcare infections, increasing hospitalization time, morbidity and mortality rates. Among several ESBLs that emerge from these pathogens, CTX-M-type enzymes had the most successful global spread in different epidemiological settings. Latin America presents high prevalence of CTX-M-2 in ESBL-producing enterobacterial infections with local emergence of the CTX-M-1 group. However, this high prevalence of the CTX-M-1 group has not yet been reported in Chile. The aim of this study was to identify ESBLs among enterobacteria isolated from clinical samples of critically ill patients from southern Chile. One-hundred thirty seven ESBL-producing bacteria were isolated from outpatients from all critical patient units from Hernán Henríquez Aravena Hospital. Phenotype characterization was performed by antibiogram, screening of ESBL, and determination of minimum inhibitory concentration (MIC). PCR was used for genetic confirmation of resistance. Molecular typing was performed by ERIC-PCR. ESBL-producing isolates were identified as Klebsiella pneumoniae (n=115), Escherichia coli (n=18), Proteus mirabilis (n=3), and Enterobacter cloacae (n=1), presenting multidrug resistance profiles. PCR amplification showed that the strains were positive for blaSHV (n=111/81%), blaCTX-M-1 (n=116/84.7%), blaTEM (n=100/73%), blaCTX-M-2 (n=28/20.4%), blaCTX-M-9 (0.7%), blaPER-1 (0.7%), and blaGES-10 (0.7%). The multiple production of ESBL was observed in 93% of isolates, suggesting high genetic mobility independent of the clonal relationship. The high frequency of the CTX-M-1 group and a high rate of ESBL co-production are changing the epidemiology of the ESBL profile in Chilean intensive care units. This epidemiology is a constant and increasing challenge, not only in Chile, but worldwide.
Assuntos
Infecções por Enterobacteriaceae/enzimologia , Infecções por Enterobacteriaceae/epidemiologia , Enterobacteriaceae/enzimologia , Unidades de Terapia Intensiva/estatística & dados numéricos , beta-Lactamases/genética , Antibacterianos/farmacologia , Chile/epidemiologia , DNA Bacteriano , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/isolamento & purificação , Infecções por Enterobacteriaceae/microbiologia , Técnicas de Genotipagem , Humanos , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Prevalência , Valores de Referência , Fatores de Risco , beta-Lactamases/isolamento & purificaçãoRESUMO
ABSTRACT Enterobacteria-producing extended-spectrum β-lactamases (ESBL) play an important role in healthcare infections, increasing hospitalization time, morbidity and mortality rates. Among several ESBLs that emerge from these pathogens, CTX-M-type enzymes had the most successful global spread in different epidemiological settings. Latin America presents high prevalence of CTX-M-2 in ESBL-producing enterobacterial infections with local emergence of the CTX-M-1 group. However, this high prevalence of the CTX-M-1 group has not yet been reported in Chile. The aim of this study was to identify ESBLs among enterobacteria isolated from clinical samples of critically ill patients from southern Chile. One-hundred thirty seven ESBL-producing bacteria were isolated from outpatients from all critical patient units from Hernán Henríquez Aravena Hospital. Phenotype characterization was performed by antibiogram, screening of ESBL, and determination of minimum inhibitory concentration (MIC). PCR was used for genetic confirmation of resistance. Molecular typing was performed by ERIC-PCR. ESBL-producing isolates were identified as Klebsiella pneumoniae (n = 115), Escherichia coli (n = 18), Proteus mirabilis (n = 3), and Enterobacter cloacae (n = 1), presenting multidrug resistance profiles. PCR amplification showed that the strains were positive for blaSHV (n = 111/81%), blaCTX-M-1 (n = 116/84.7%), blaTEM (n = 100/73%), blaCTX-M-2 (n = 28/20.4%), blaCTX-M-9 (0.7%), blaPER-1 (0.7%), and blaGES-10 (0.7%). The multiple production of ESBL was observed in 93% of isolates, suggesting high genetic mobility independent of the clonal relationship. The high frequency of the CTX-M-1 group and a high rate of ESBL co-production are changing the epidemiology of the ESBL profile in Chilean intensive care units. This epidemiology is a constant and increasing challenge, not only in Chile, but worldwide.
Assuntos
Humanos , beta-Lactamases/genética , Enterobacteriaceae/enzimologia , Infecções por Enterobacteriaceae/enzimologia , Infecções por Enterobacteriaceae/epidemiologia , Unidades de Terapia Intensiva/estatística & dados numéricos , Valores de Referência , beta-Lactamases/isolamento & purificação , DNA Bacteriano , Testes de Sensibilidade Microbiana , Chile/epidemiologia , Reação em Cadeia da Polimerase , Prevalência , Fatores de Risco , Enterobacteriaceae/isolamento & purificação , Enterobacteriaceae/efeitos dos fármacos , Infecções por Enterobacteriaceae/microbiologia , Técnicas de Genotipagem , Antibacterianos/farmacologiaRESUMO
The increase in carbapenem-resistant gram-negative bacteria is a matter of concern due to the limited therapeutic options available. In severe infections caused by these isolates, the rapid detection of the mechanisms of resistance is vital. We described a slightly modified version of the Blue-Carba test, rapid Blue-Carba test, which allows the detection of carbapenemases at 4 h of incubation from a haze of bacterial growth obtained from a positive blood culture. It was able to detect carbapenemase-producing isolates (Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter baumannii) with a sensitivity and specificity of 98.1 and 100%, respectively. It is a rapid, easy-to-perform and an inexpensive technique that can be applied to routine laboratories, together with the simultaneous identification by mass spectrometry which would help to screen non-enzymatic carbapenem resistance; this method allows the detection of clinically relevant multidrug-resistant bacteria and the early implementation of accurate therapeutic interventions.
Assuntos
Infecções por Acinetobacter/enzimologia , Bacteriemia/enzimologia , Proteínas de Bactérias/sangue , Farmacorresistência Bacteriana Múltipla , Infecções por Enterobacteriaceae/enzimologia , Infecções por Pseudomonas/enzimologia , Resistência beta-Lactâmica , beta-Lactamases/sangue , Infecções por Acinetobacter/diagnóstico , Infecções por Acinetobacter/tratamento farmacológico , Infecções por Acinetobacter/microbiologia , Acinetobacter baumannii/efeitos dos fármacos , Acinetobacter baumannii/enzimologia , Acinetobacter baumannii/crescimento & desenvolvimento , Acinetobacter baumannii/isolamento & purificação , Antibacterianos/metabolismo , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Argentina , Bacteriemia/diagnóstico , Bacteriemia/tratamento farmacológico , Bacteriemia/microbiologia , Proteínas de Bactérias/genética , Carbapenêmicos/metabolismo , Carbapenêmicos/farmacologia , Carbapenêmicos/uso terapêutico , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/enzimologia , Enterobacteriaceae/crescimento & desenvolvimento , Enterobacteriaceae/isolamento & purificação , Infecções por Enterobacteriaceae/diagnóstico , Infecções por Enterobacteriaceae/tratamento farmacológico , Infecções por Enterobacteriaceae/microbiologia , Hospitais Universitários , Humanos , Inativação Metabólica , Tipagem Molecular , Infecções por Pseudomonas/diagnóstico , Infecções por Pseudomonas/tratamento farmacológico , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/enzimologia , Pseudomonas aeruginosa/crescimento & desenvolvimento , Pseudomonas aeruginosa/isolamento & purificação , Sensibilidade e Especificidade , Fatores de Tempo , beta-Lactamases/genéticaRESUMO
Carbapenem-resistant Enterobacteriaceae carrying New Delhi metallo-ß-lactamase 1 (NDM-1) have rarely been reported in Latin America. We report of an outbreak caused by a blaNDM-1-harboring plasmid spread through different bacterial species, including Escherichia coli (ST617) and Enterobacter cloacae (ST182) isolates from the same patient and three Klebsiella pneumoniae isolates (ST22) derived from three epidemiologically related patients. IncFII plasmids were found in all strains. Measures to control the outbreak were applied successfully.
Assuntos
Enterobacteriaceae/enzimologia , Enterobacteriaceae/fisiologia , Plasmídeos/genética , beta-Lactamases/metabolismo , Antibacterianos/uso terapêutico , Surtos de Doenças , Enterobacteriaceae/efeitos dos fármacos , Infecções por Enterobacteriaceae/tratamento farmacológico , Infecções por Enterobacteriaceae/enzimologia , Feminino , Genótipo , Humanos , Masculino , México , Atenção Terciária à Saúde , beta-Lactamases/genéticaRESUMO
OBJECTIVE: To analyze the profile of patients with microorganisms resistant to carbapenems, and the prevalence of the enzyme Klebsiella pneumoniae carbapenemase in interobacteriaceae. METHODS: Retrospective descriptive study. From the isolation in bacteriological tests ordered by clinicians, we described the clinical and epidemiological characteristics of patients with enterobacteria resistants to carbapenems at a university hospital, between March and October 2013. RESULTS: We included 47 isolated patients in this study, all exhibiting resistance to carbapenems, including 9 patients who were confirmed as infected/colonized with K. pneumoniae carbapenemase. Isolation in tracheal aspirates (12; 25.5%) predominated. The resistance to ertapenem, meropenem, and imipenem was 91.5%, 83.0% and 80.0%, respectively. Aminoglycosides was the class of antimicrobials that showed the highest sensitivity, 91.5% being sensitive to amikacin and 57.4% to gentamicin. CONCLUSION: The K. pneumoniae carbapenemase was an important agent in graun isotaling in hospital intection. The limited therapeutic options emphasize the need for rapid laboratory detection, as well as the implementation of measures to prevent and control the spread of these pathogens.
Assuntos
Proteínas de Bactérias/biossíntese , Infecção Hospitalar/microbiologia , Farmacorresistência Bacteriana , Enterobacteriaceae/enzimologia , Infecções por Klebsiella/enzimologia , Klebsiella pneumoniae/enzimologia , beta-Lactamases/biossíntese , Adolescente , Adulto , Carbapenêmicos/uso terapêutico , Criança , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/isolamento & purificação , Infecções por Enterobacteriaceae/enzimologia , Infecções por Enterobacteriaceae/microbiologia , Feminino , Hospitais de Ensino/estatística & dados numéricos , Humanos , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/isolamento & purificação , Masculino , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Estudos Retrospectivos , Adulto JovemRESUMO
Objective To analyze the profile of patients with microorganisms resistant to carbapenems, and the prevalence of the enzyme Klebsiella pneumoniae carbapenemase in interobacteriaceae. Methods Retrospective descriptive study. From the isolation in bacteriological tests ordered by clinicians, we described the clinical and epidemiological characteristics of patients with enterobacteria resistants to carbapenems at a university hospital, between March and October 2013. Results We included 47 isolated patients in this study, all exhibiting resistance to carbapenems, including 9 patients who were confirmed as infected/colonized with K. pneumoniae carbapenemase. Isolation in tracheal aspirates (12; 25.5%) predominated. The resistance to ertapenem, meropenem, and imipenem was 91.5%, 83.0% and 80.0%, respectively. Aminoglycosides was the class of antimicrobials that showed the highest sensitivity, 91.5% being sensitive to amikacin and 57.4% to gentamicin. Conclusion The K. pneumoniae carbapenemase was an important agent in graun isotaling in hospital intection. The limited therapeutic options emphasize the need for rapid laboratory detection, as well as the implementation of measures to prevent and control the spread of these pathogens. .
Objetivo Analisar o perfil dos pacientes que apresentaram microrganismos com resistência aos carbapenêmicos, e a prevalência da enzima Klebsiella pneumoniae carbapenemase em enterobactérias. Métodos Estudo retrospectivo descritivo. A partir do isolamento em exames bacteriológicos solicitados pelos clínicos, descrevemos as características clínicas e epidemiológicas dos pacientes que apresentaram enterobactérias resistentes aos carbapenêmicos entre março e outubro de 2013 em um hospital universitário. Resultados Foram incluídos 47 pacientes isolados, todos apresentando resistência aos carbapenêmicos, dos quais 9 tiveram confirmação de infecção/colonização por K. pneumoniae carbapenemase. Ocorreu predomínio de isolamento em aspirados traqueais (12; 25,5%). A resistência ao ertapenem, meropenem e imipenem foi de 91,5%, 83,0% e 80,0%, respectivamente. Os aminoglicosídeos foram a classe de antimicrobianos que apresentou maior sensibilidade, sendo 91,5% sensível a amicacina e 57,4% a gentamicina. Conclusão A K. pneumoniae carbapenemase constituiu um importante patógeno hospitalar em isolamento crescente nesse nosocômio. As limitadas opções terapêuticas reforçam a necessidade de uma rápida detecção laboratorial, assim como a implementação de medidas de prevenção e controle da disseminação desses patógenos. .
Assuntos
Adolescente , Adulto , Criança , Feminino , Humanos , Masculino , Adulto Jovem , Proteínas de Bactérias/biossíntese , Infecção Hospitalar/microbiologia , Farmacorresistência Bacteriana , Enterobacteriaceae/enzimologia , Infecções por Klebsiella/enzimologia , Klebsiella pneumoniae/enzimologia , beta-Lactamases/biossíntese , Carbapenêmicos/uso terapêutico , Infecções por Enterobacteriaceae/enzimologia , Infecções por Enterobacteriaceae/microbiologia , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/isolamento & purificação , Hospitais de Ensino/estatística & dados numéricos , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/isolamento & purificação , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Estudos RetrospectivosAssuntos
Carbapenêmicos/farmacologia , Resistência a Múltiplos Medicamentos , Infecções por Enterobacteriaceae/enzimologia , Providencia/efeitos dos fármacos , Providencia/enzimologia , beta-Lactamases/efeitos dos fármacos , Antibacterianos/farmacologia , Suscetibilidade a Doenças , Ertapenem , Humanos , Meropeném , Tienamicinas/farmacologia , beta-Lactamases/biossíntese , beta-Lactamas/farmacologiaRESUMO
The emergence of Extended-Spectrum Beta-Lactamase (ESBL)-producing microorganisms in Brazilian hospitals is a challenge that concerns scientists, clinicians and healthcare institutions due to the serious risk they pose to confined patients. The goal of this study was the detection of ESBL production by clinical strains of Escherichia coli and Klebsiella sp. isolated from pus, urine and blood of patients at Hospital Universitário Santa Maria, Rio Grande Sul, RS, Brazil and the genotyping of the isolates based on bla SHV genes. The ESBL study was carried out using the Combined Disc Method, while Polymerase Chain Reaction (PCR) was used to study the bla SHV genes. Of the 90 tested isolates, 55 (61.1 percent) were identified as ESBL-producing by the combined disk method. The bla SHV genes were found in 67.8 percent of these microorganisms. K. pneumoniae predominated in the samples, presenting the highest frequency of positive results from the combined disk and PCR.
Assuntos
Humanos , Sequência de Bases , Ensaios Enzimáticos Clínicos , Resistência Microbiana a Medicamentos , Enterobacteriaceae/enzimologia , Infecções por Enterobacteriaceae/enzimologia , Pacientes , Reação em Cadeia da Polimerase , Pseudomonadaceae/enzimologia , beta-Lactamases/análise , Técnicas e Procedimentos Diagnósticos , Genótipo , Métodos , PrevalênciaRESUMO
Analysis of zymograms with SDS-polyacrilamide gel electrophoresis containing gelatin as substrate, and performed on samples of haemolymph or fat body taken from Rhodnius prolixus inoculated or not with Enterobacter cloacae, demonstrated distinct patterns of protease activities: (i) in the haemolymph two proteases were induced in insects inoculated with bacteria; (ii) two proteases were detected in the fat bodies derived from non-inoculated controls or insect inoculated with sterile culture medium; (iii) haemolymph and fat body had both the same apparent molecular weights proteases (46 and 56 kDa); and (iv) these enzymes were characterized as metallo-proteases. The association of these enzymes in Rhodnius infected with bacteria was discussed.