Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 35
Filtrar
1.
Int J Antimicrob Agents ; 64(2): 107229, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38823493

RESUMO

OBJECTIVES: Therapeutic drug monitoring (TDM) of ß-lactam antibiotics in critically ill patients may benefit dose optimisation, thus improving therapeutic outcomes. However, rapidly and accurately detecting these antibiotics in blood remains a challenge. This research group recently developed a thermometric biosensor called the New Delhi metallo-ß-lactamase-1 (NDM-1) biosensor, which detects multiple classes of ß-lactam antibiotics in spiked plasma samples. METHODS: This study assessed the NDM-1 biosensor's effectiveness in detecting plasma concentrations of ß-lactam antibiotics in treated patients. Seven patients receiving cefuroxime were studied. Plasma samples collected pre- and post-antibiotic treatment were analysed using the NDM-1 biosensor and compared with liquid chromatography coupled with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). RESULTS: The biosensor detected plasma samples without dilution, and a brief pre-treatment using a polyvinylidene fluoride filter significantly lowered matrix effects, reducing the running time to 5-8 minutes per sample. The assay's linear range for cefuroxime (6.25-200 mg/L) covered target concentrations during the trough phase of pharmacokinetics in critically ill patients. The pharmacokinetic properties of cefuroxime in treated patients determined by the NDM-1 biosensor and the UPLC-MS/MS were comparable, and the cefuroxime plasma concentrations measured by the two methods showed statistically good consistency. CONCLUSION: These data demonstrate that the NDM-1 biosensor assay is a fast, sensitive, and accurate method for detecting cefuroxime plasma concentrations in treated patients and highlights the NDM-1 biosensor as a promising tool for on-site TDM of ß-lactam antibiotics in critically ill patients.


Assuntos
Antibacterianos , Técnicas Biossensoriais , Cefuroxima , Monitoramento de Medicamentos , Espectrometria de Massas em Tandem , beta-Lactamases , Humanos , beta-Lactamases/sangue , Antibacterianos/uso terapêutico , Antibacterianos/sangue , Cefuroxima/sangue , Cefuroxima/uso terapêutico , Monitoramento de Medicamentos/métodos , Técnicas Biossensoriais/métodos , Masculino , Pessoa de Meia-Idade , Feminino , Idoso , Cromatografia Líquida/métodos , Plasma/química , Estado Terminal
3.
J Glob Antimicrob Resist ; 19: 81-82, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31493526

RESUMO

OBJECTIVES: Klebsiella pneumoniae is a well-known pathogen frequently implicated in serious life-threatening nosocomial infections. Here we present a K. pneumoniae isolate (AHEPA1046) co-harbouring blaNDM-1 and blaOXA-48 isolated from a blood sample of an inpatient in Thessaloniki, Greece. METHODS: Whole-genome sequencing (WGS) was performed using an Illumina MiniSeq Sequencing System. Multilocus sequence typing (MLST) was performed using a BLAST-based approach, and antimicrobial resistance genes and plasmid replicons were identified by ResFinder and PlasmidFinder, respectively. The Rapid Annotation using Subsystem Technology (RAST) v.2.0 server was used for genome annotation. RESULTS: WGS analysis revealed the complete resistome of K. pneumoniae AHEPA1046. The strain harboured blaNDM-1 and blaOXA-48 together with 16 additional antimicrobial resistance genes and was resistant to carbapenems, aminoglycosides, quinolones, macrolides, tetracyclines, trimethoprim, fosfomycin and phenicols. Moreover, it was classified as ST11. CONCLUSION: This is the first report of a K. pneumoniae clinical isolate from Greece co-producing NDM-1 and OXA-48 carbapenemases and is one of a few reported worldwide.


Assuntos
Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Farmacorresistência Bacteriana Múltipla/genética , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/metabolismo , beta-Lactamases/genética , beta-Lactamases/metabolismo , Adulto , Antibacterianos/farmacologia , Proteínas de Bactérias/sangue , Composição de Bases , Sequência de Bases , Genes Bacterianos/genética , Grécia , Humanos , Masculino , Tipagem de Sequências Multilocus , Plasmídeos , Sequenciamento Completo do Genoma , beta-Lactamases/sangue
4.
Braz J Microbiol ; 50(3): 657-662, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31270693

RESUMO

The emergence of carbapenem-resistant Enterobacterales (CRE) is a matter of public health concern. Carbapenemases are the main mechanism of resistance among CRE, and its rapid detection is essential. The detection of carbapenemases usually requires culture-based methods and molecular assays, which may be costly and need long turnaround times. Recently, an easy and rapid immunochromatographic assay for carbapenemases (OXA-48, KPC, and NDM) detection based in lateral flow immunoassay with specific monoclonal antibodies on a nitrocellulose membrane has been developed. We aimed to evaluate the RESIST-3 O.K.N. in colonies from pure culture as well as in spiked blood cultures with Enterobacterales. All carbapenemase producers (CP) presenting the OXA-48-like, KPC, and NDM enzymes presented positive results in both pure colonies and spiked blood cultures. None of the carbapenemase non-producers (CNP) presented positive results in the tests. A total of 97% CP isolates presented positive results in pure colonies in less than 5 min. For CP directly from blood culture, the mean time to positivity for OXA-48-like and KPC was 1 min, whereas it was 25 min for NDM. Our results indicate that this immunoassay can be used to detect carbapenemases directly from blood culture bottles in a routine diagnostic laboratory, which would reduce the turnaround time of CP detection.


Assuntos
Proteínas de Bactérias/sangue , Infecções por Enterobacteriaceae/sangue , Enterobacteriaceae/enzimologia , Imunoensaio/métodos , beta-Lactamases/sangue , Antibacterianos/farmacologia , Hemocultura , Brasil , Carbapenêmicos/farmacologia , Farmacorresistência Bacteriana , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/genética , Infecções por Enterobacteriaceae/diagnóstico , Infecções por Enterobacteriaceae/microbiologia , Humanos
6.
J Clin Microbiol ; 56(9)2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29950338

RESUMO

The aim of this study was to evaluate the two rapid colorimetric methods (CNPt-Direct and Blue-Carba) for the detection of carbapenemase production directly from blood culture in a routine microbiology laboratory. The methods were initially evaluated on spiked blood cultures with 61 carbapenemase-positive isolates. Afterwards, they were used in blood cultures (314 samples were evaluated) obtained from patients in a routine microbiology laboratory during a period of 6 months. The colorimetric methods were compared to the conventional culture of blood. The results of the spiked blood cultures indicated that both colorimetric methods presented positive results for the vast majority (95%) of the isolates harboring KPC, NDM, and IMP genes. However, the assay failed to detect many GES- and OXA-48-like-positive isolates (65% positive results). In the second part of the study, a total of 314 blood cultures from patients were evaluated, and 33 yielded Enterobacteriaceae isolates resistant to meropenem (30 isolates were positive for carbapenemases according to PCR). The colorimetric tests correctly detected 24 out of the 30 carbapenemase-positive isolates directly from the blood vial (80% positive results). Overall positive percent agreement and negative percent agreement were 80% and 100%, respectively. The colorimetric assays are simple and cost-effective methods that can be implemented in a routine microbiology laboratory, diminishing the time necessary to detect carbapenemase-producing isolates from 24 to 48 h to 3 to 5 h. Moreover, according to our results, the positive colorimetric test results do not need to be confirmed and can be immediately provided to the attending physician.


Assuntos
Proteínas de Bactérias/sangue , Técnicas Bacteriológicas/métodos , Hemocultura/métodos , Colorimetria/métodos , Testes Diagnósticos de Rotina/métodos , beta-Lactamases/sangue , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Hemocultura/normas , Farmacorresistência Bacteriana , Enterobacteriaceae/enzimologia , Enterobacteriaceae/genética , Infecções por Enterobacteriaceae/microbiologia , Reações Falso-Negativas , Humanos , Meropeném/farmacologia , Kit de Reagentes para Diagnóstico , Sensibilidade e Especificidade , beta-Lactamases/genética
7.
Aging Clin Exp Res ; 30(7): 839-843, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29032522

RESUMO

BACKGROUND AND AIMS: To examine antimicrobial resistance of commonly isolated pathogens in elderly hospitalized patients. METHODS: Data regarding all clinically significant isolates from blood and urine cultures of patients admitted to a multilevel geriatric hospital during March 2015 to April 2016 were collected. Antimicrobial susceptibility testing was performed according to Clinical and Laboratory Standard Institute guidelines. RESULTS: Escherichia coli, Proteus mirabilis, and Klebsiella pneumoniae were the most common isolates, with proportions of extended spectrum beta-lactamase positivity of 60, 40, and 61% respectively. Adjusted logistic regression models indicated that resistance of Escherichia coli to ceftriaxone [odds ratio (OR) 2.8, 95% confidence interval (CI) 1.5-5.1], ceftazidime (OR 2.8, 95% CI 1.5-5.1), ciprofloxacin (OR 2.2, 95% CI 1.2-4.0), amoxicillin/clavulanic acid (OR 2.3, 95% CI 1.2-4.3), and trimethoprim/sulfamethoxazole (OR 2.4, 95% CI 1.4-4.3) was significantly higher in skilled nursing wards than in acute geriatric wards. Resistance of Proteus mirabilis to ceftriaxone (OR 3.1, 95% CI 1.5-6.4) and Klebsiella pneumoniae to ciprofloxacin (OR 3.2, 95% CI 1.3-7.9) was significantly higher in skilled nursing wards than in acute wards. CONCLUSIONS AND DISCUSSION: Antimicrobial resistance was found to be high in a multilevel geriatric hospital, especially in skilled nursing wards. These findings call for rethinking of the empirical antimicrobial therapy and of the efforts for prevention of nosocomial infection.


Assuntos
Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana/fisiologia , Escherichia coli/efeitos dos fármacos , Klebsiella pneumoniae/efeitos dos fármacos , Proteus mirabilis/efeitos dos fármacos , Idoso , Antibacterianos/farmacologia , Infecção Hospitalar/prevenção & controle , Escherichia coli/isolamento & purificação , Feminino , Geriatria , Hospitais Especializados , Humanos , Klebsiella pneumoniae/isolamento & purificação , Masculino , Proteus mirabilis/isolamento & purificação , Estudos Retrospectivos , beta-Lactamases/sangue , beta-Lactamases/urina
8.
J Chemother ; 29(3): 150-153, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27347714

RESUMO

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/genética
10.
Biomed Res Int ; 2016: 4307987, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26904674

RESUMO

Recently, Wong et al. have successfully developed a fluorescent biosensor based on the PenPC ß-lactamase which changes its intrinsic fluorescence in presence of ß-lactam antibiotics (BLAs). Here, we studied systematically this correlation among the fluorescence change of the biosensor and the concentration of different BLAs aimed at developing a novel method for estimating the concentration of a wide range of BLAs. This method showed high precision and specificity and very low interference from clinically relevant samples. We were able to monitor the pharmacokinetics of meropenem in healthy volunteers as well as in an ill animal model too, indicating that the implemented method could be suitable for clinical practice.


Assuntos
Antibacterianos/sangue , Técnicas Biossensoriais/métodos , Tienamicinas/sangue , beta-Lactamas/sangue , Animais , Antibacterianos/farmacocinética , Antibacterianos/uso terapêutico , Monitoramento de Medicamentos , Fluorescência , Voluntários Saudáveis , Humanos , Meropeném , Sus scrofa/microbiologia , Tienamicinas/farmacocinética , Tienamicinas/uso terapêutico , beta-Lactamases/sangue , beta-Lactamas/farmacocinética , beta-Lactamas/uso terapêutico
11.
J Infect Chemother ; 21(10): 742-6, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26143049

RESUMO

INTRODUCTION: We sought to describe a case of pharmacodynamically-optimized dosing of piperacillin-tazobactam in a patient that cleared their infections after treatment with high-dose, extended-infusion piperacillin-tazobactam and summarize the literature on the benefits of extended-infusion of beta-lactams. CASE REPORT: At an outside hospital, a 78 year-old male presented with fevers and shortness of breath. He was empirically initiated on standard doses of vancomycin and piperacillin-tazobactam for suspected pneumonia and sepsis. Blood and sputum cultures identified Elizabethkingia meningosepticum sensitive only to piperacillin-tazobactam by E-test susceptibility testing. After 10 days of empiric therapy with piperacillin-tazobactam dosed at 3.375 g IV every 8 h over 30 min, the patient transferred to our institution and was initiated on piperacillin-tazobactam at 3.375 g IV every 8 h administered as a 4 h infusion. The patient failed to improve; piperacillin-tazobactam was changed to 4.5 g IV over 4 h every 8 h and later changed to the hospital protocol dose of 3.375 g IV over 4 h every 6 h. The patient achieved negative blood cultures within 24 h of optimized dosing. DISCUSSION: We present the first case to our knowledge that describes failure to respond and subsequent response within a single patient where beta-lactam dosing was altered to optimize pharmacokinetics and pharmacodynamics (PK-PD). Our patient received non-standard dose-escalation for piperacillin-tazobactam. Drug exposure was estimated post-hoc utilizing robust mathematical simulations to describe alterations in disposition over time. This case demonstrates that extended-infusion administration of beta-lactams may provide improved microbiological activity.


Assuntos
Antibacterianos/administração & dosagem , Bacteriemia/tratamento farmacológico , Endocardite/terapia , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Ácido Penicilânico/análogos & derivados , beta-Lactamases/sangue , Idoso , Bacteriemia/etiologia , Endocardite/etiologia , Infecções por Bactérias Gram-Negativas/complicações , Humanos , Infusões Intravenosas , Masculino , Ácido Penicilânico/administração & dosagem , Piperacilina/administração & dosagem , Combinação Piperacilina e Tazobactam , beta-Lactamas/uso terapêutico
12.
J Antimicrob Chemother ; 69(8): 2132-6, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24722840

RESUMO

OBJECTIVES: Recently, matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) was successfully applied for the detection of carbapenemase activity directly from Gram-negative colonies. Based on this principle, we evaluated the performance of MALDI-TOF MS for rapid detection of carbapenemase activity directly from positive blood culture vials. METHODS: A total of 100 blood culture vials were randomly selected. MALDI-TOF MS carbapenemase assay results were confirmed by the detection of carbapenemase-encoding genes. RESULTS: A total of 110 bacterial isolates were recovered. The MALDI-TOF MS carbapenemase assay identified 21 of 29 (72.4%) of the carbapenemase-producing isolates directly from the blood culture vials, especially those encoding KPC-2 (100%) and SPM-1 (100%), after a 4 h incubation period. Although the majority of OXA-23-producing Acinetobacter baumannii isolates were not identified on day 1, all isolates were identified as carbapenemase producers directly from the colony on the next day. CONCLUSIONS: The MALDI-TOF MS carbapenemase assay is a feasible and rapid test to identify carbapenemase activity directly from blood culture vials. It may contribute to faster readjustment of empirical antimicrobial therapy and implementation of infection control measures.


Assuntos
Proteínas de Bactérias/sangue , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , beta-Lactamases/sangue , Acinetobacter baumannii/enzimologia , Acinetobacter baumannii/isolamento & purificação , Humanos , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/isolamento & purificação , Pseudomonas aeruginosa/enzimologia , Pseudomonas aeruginosa/isolamento & purificação
13.
J Exp Med ; 211(1): 153-63, 2014 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-24367002

RESUMO

Signal transducer and activator of transcription 5 (Stat5) is essential for cytokine-regulated processes such as proliferation, differentiation, and survival in hematopoietic cells. To investigate the role of Stat5 in osteoclasts, we generated mice with an osteoclast-specific conditional deletion of Stat5 (Stat5 conditional knockout [cKO] mice) and analyzed their bone phenotype. Stat5 cKO mice exhibited osteoporosis caused by an increased bone-resorbing activity of osteoclasts. The activity of mitogen-activated protein kinases (MAPKs), in particular extracellular signal-related kinase, was increased in Stat5 cKO osteoclasts, whereas the expression of the MAPK phosphatases dual specificity phosphatase 1 (Dusp1) and Dusp2 was significantly decreased. Interleukin-3 (IL-3) stimulated the phosphorylation and nuclear translocation of Stat5 in osteoclasts, and Stat5 expression was up-regulated in response to receptor activator of nuclear factor κB ligand (RANKL). The results suggest that Stat5 negatively regulates the bone-resorbing function of osteoclasts by promoting Dusp1 and Dusp2 expression, and IL-3 promotes Stat5 activation in osteoclasts.


Assuntos
Reabsorção Óssea/fisiopatologia , Fosfatase 1 de Especificidade Dupla/metabolismo , Retroalimentação Fisiológica/fisiologia , Regulação Enzimológica da Expressão Gênica/fisiologia , Osteoclastos/fisiologia , Fator de Transcrição STAT5/metabolismo , Animais , Western Blotting , Densidade Óssea , Reabsorção Óssea/genética , Primers do DNA/genética , Regulação Enzimológica da Expressão Gênica/genética , Técnicas Histológicas , Marcação In Situ das Extremidades Cortadas , Fator de Crescimento Insulin-Like I/metabolismo , Interleucina-3/metabolismo , Camundongos , Camundongos Knockout , Análise em Microsséries , Osteocalcina/sangue , Reação em Cadeia da Polimerase , Reação em Cadeia da Polimerase em Tempo Real , Fator de Transcrição STAT5/genética , beta-Lactamases/sangue
14.
Adv Clin Exp Med ; 23(6): 865-70, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25618110

RESUMO

BACKGROUND: The prevalence of bloodstream infections (BSIs) due to ESBL-producing Escherichia coli (ESBL-EC) strains has increased dramatically over the past years. OBJECTIVES: Characterization of ESBL-EC isolates collected from BSIs with regard to their antimicrobial susceptibility and phylogenetic background. The conjugative transfer of resistance determinants to the E. coli reference strain K12 C600 was also investigated. MATERIAL AND METHODS: A collection of forty-eight ESBL-EC strains recovered from BSIs was subjected to the study. These strains were obtained from the ICU (intensive care unit) of the Medical University Hospital, Wroclaw, Poland, during a four-year period (2009-2012). All the isolates were screened for ESBL production by the double disk synergy test (DDST). Transferability of plasmid-mediated resistance genes was performed by the conjugational broth method. Susceptibility to antibiotics and chemotherapeutics of clinical isolates and transconjugants was determined by the agar dilution method. PCR assay was used to detect the blaCTX-M gene in ESBL-EC tested and transconjugants. Affiliation to phylogenetic groups was done by the triplex PCR method. RESULTS: Conjugational transfer of plasmids responsible for ESBL to a recipient strain was successful for all the ESBL-EC analyzed (donors). The conjugation frequencies ranging from 2.3×10(-7) to 5.2×10(-1) per donor. In vitro susceptibility testing revealed that all the ESBL-EC isolates and their transconjugants were resistant to most of the antimicrobial agents tested with the exception of carbapenems, tigecycline, and ß-lactam-clavulanate combinations. Moreover, all the donor strains and their transconjugants were found to contain the blaCTX-M gene. The majority of the isolates analyzed belonged to phylogroups B2 (62.5%) and D (25%), whereas groups B1 and A were less frequently represented (8.3% and 4.2%, respectively). CONCLUSIONS: The results of the study confirm the need of antibiotic policies and effective infection control measures in hospital settings to minimize BSIs caused by multi-resistant ESBL-producing pathogens.


Assuntos
Bacteriemia/tratamento farmacológico , Infecção Hospitalar/tratamento farmacológico , Infecções por Escherichia coli/tratamento farmacológico , Escherichia coli/efeitos dos fármacos , Filogenia , Resistência beta-Lactâmica , beta-Lactamases/sangue , beta-Lactamas/uso terapêutico , Bacteriemia/sangue , Bacteriemia/diagnóstico , Bacteriemia/microbiologia , Bacteriemia/transmissão , Infecção Hospitalar/diagnóstico , Infecção Hospitalar/microbiologia , Infecção Hospitalar/transmissão , Farmacorresistência Bacteriana Múltipla , Escherichia coli/enzimologia , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/sangue , Infecções por Escherichia coli/diagnóstico , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/transmissão , Feminino , Genótipo , Hospitais Universitários , Humanos , Unidades de Terapia Intensiva , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Fenótipo , Polônia , Reação em Cadeia da Polimerase , Fatores de Tempo , Resistência beta-Lactâmica/genética , beta-Lactamases/genética
15.
Acta bioquím. clín. latinoam ; Acta bioquím. clín. latinoam;46(3): 405-412, set. 2012. ilus, graf, tab
Artigo em Espanhol | LILACS | ID: lil-662034

RESUMO

La rápida emergencia de resistencia a antimicrobianos debida a la presencia de b-lactamasas de espectro extendido (BLEE) tiene un impacto significativo en la salud pública. Las BLEEs son enzimas producidas por bacilos gramnegativos y confieren resistencia a las penicilinas, a todas las cefalosporinas y al aztreonam, pero no a los carbapenemes ni a las cefamicinas y la mayoría son inhibidas por el ácido clavulánico. El objetivo de este trabajo fue evaluar la resistencia a antibióticos b-lactámicos en aislamientos de Klebsiella pneumoniae, Escherichia coli y Proteus mirabilis y caracterizar las b-lactamasas responsables de dicha resistencia. Se analizaron 2.030 aislamientos (362 Klebsiella pneumoniae, 1.250 Escherichia coli y 175 Proteus mirabilis) provenientes de diferentes materiales clínicos de pacientes que concurrieron al Hospital Provincial del Centenario de la ciudad de Rosario (Santa Fe) durante el período 2008-2009. Los ensayos de sensibilidad antibiótica se realizaron de acuerdo con las recomendaciones del Clinical and Laboratory Standard Institute. Se confirmó la presencia de los genes codificantes de BLEE blaTEM, blaSHV, blaCTX-M y blaPER mediante la reacción en cadena de la polimerasa (PCR) utilizando cebadores específicos. Los aislados fueron caracterizados fenotípicamente como productores de BLEE y demostraron poseer varios genes bla. Se detectaron tres diferentes b-lactamasas BLEE derivadas de SHV, TEM y CTX-M y se demostró que pueden coexistir dos o más de estos genes en una misma bacteria.


The rapid emergence of antimicrobial resistance due to extended spectrum b-lactamases (ESBL) has a significant impact on public health. ESBL, produced by gram-negative bacilli, are enzymes that confer resistance to penicillins, cephalosporins and aztreonam, but not to carbapenems or cephamycins, and are usually inhibited by clavulanic acid. The aim of this study was to evaluate b-lactam resistance within isolates of Klebsiella pneumoniae, Escherichia coli, and Proteus mirabilis and to characterize the b-lactamases responsible for this resistance. A total of 2,030 strains (362 Klebsiella pneumoniae, 1,250 Escherichia coli, and 175 Proteus mirabilis) isolated from patients at Hospital Provincial del Centenario in Rosario-Santa Fe were analyzed from 2008 to 2009. Antibiotic sensitivity tests were performed according to Clinical and Laboratory Standard Institute recommendations. Molecular detection of ESBL-related bla genes, including blaTEM, blaSHV, blaCTX-M and blaPER was performed by polymerase chain reaction (PCR) using specific primers. The strains were phenotipically confirmed as ESBL producers and the isolates carried several bla genes. Three different b-lactamases were detected: SHV-related, TEM-related and CTX-M-related, showing that two or more genes may coexist in the same bacterium.


A rápida emergência de resistência a antimicrobianos devida à presença de b lactamases de espectro estendido (BLEE) tem um impacto significativo na saúde pública. As BLEEs são enzimas produzidas por bacilos gram-negativos e conferem resistência às penicilinas, a todas as cefalosporinas e ao aztreonam, mas não aos carbapenêmicos nem às cefamicinas e a maioria são inibidas pelo ácido clavulânico. O objetivo deste trabalho foi avaliar a resistência a antibióticos b-lactâmicos em isolamentos de Klebsiella pneumoniae, Escherichia coli e Proteus mirabilis e caracterizar as b-lactamases responsáveis por tal resistência. Foram analisados 2.030 isolamentos (362 Klebsiella pneumoniae, 1.250 Escherichia coli e 175 Proteus mirabilis) provenientes de diferentes materiais clínicos de pacientes que foram ao Hospital Provincial do Centenário da cidade de Rosario (Santa Fe) durante o período 2008-2009. Os ensaios de sensibilidade antibiótica foram realizados de acordo com as recomendações do Clinical and Laboratory Standard Institute. Confirmou-se a presença dos genes codificantes de BLEE blaTEM, blaSHV, blaCTX-M e blaPER mediante a reação em cadeia da polimerase (PCR) utilizando cevadores específicos. Os isolados foram caracterizados fenotipicamente como produtores de BLEE e demonstraram possuir vários genes bla. Foram detectadas três diferentes b-lactamases derivadas de SHV, TEM e CTX-M e se demonstrou que podem coexistir dois ou mais destes genes numa mesma bactéria.


Assuntos
Humanos , Inibidores de beta-Lactamases/sangue , Inibidores de beta-Lactamases/urina , beta-Lactamases/sangue , Argentina , Resistência beta-Lactâmica , Resistência Microbiana a Medicamentos , Escherichia coli , Klebsiella pneumoniae , Proteus mirabilis
16.
Wien Klin Wochenschr ; 124(15-16): 504-15, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22895677

RESUMO

BACKGROUND AND AIM: The aim of the study was to characterize ESBL-producing uropathogenic Escherichia coli (UPEC) strains isolated in children. That included the investigation of virulence factors and the analysis of the types of ß-lactamases at the molecular genetic level. MATERIAL AND METHODS: During the 2-year study period, 77 ESBL-producing E. coli strains were recovered from urine samples of febrile children with significant bacteriuria hospitalized at one Croatian hospital. Susceptibility of isolates to bactericidal serum activity was tested by Shiller and Hatch method, while adhesin expression was determined by agglutination methods. Characterization of ESBLs was performed by PCR with specific primers for ESBLs and by sequencing of bla (ESBL) genes. Genotyping of the E. coli isolates was performed by pulsed-field gel electrophoresis (PFGE). RESULTS: Twenty-seven (35.1 %) and 50 (64.9 %) ESBL-producing UPEC strains were isolated in neonates and infants, respectively. Of 70 strains investigated for the presence of virulence factors, adhesins were detected in 48.6 % strains (8.6 % in the neonate and 40 % in the infants group) giving a statistically significant difference in adhesin expression between the two groups (p < 0.01). Hemolysin was produced by 84.3 %, whereas 70 % of strains were serum-resistant. The bla (TEM) gene was detected in 22 (28 %) and bla (SHV) gene in 57 strains (74 %), whereas bla (CTX-M) gene was detected in only two isolates (2.5%). In ten isolates, bla (TEM) and bla (SHV) were simultaneously detected. Sequencing of bla (SHV) genes revealed that SHV-5 ß-lactamase was by far the most prevalent and was found in 51 strains (66 %). The strains were clonally related as demonstrated by PFGE and assigned into ten clusters. CONCLUSIONS: Infection control measures should be employed and the consumption of expanded-spectrum cephalosporins in the hospital should be restricted.


Assuntos
Escherichia coli Uropatogênica/enzimologia , Escherichia coli Uropatogênica/isolamento & purificação , beta-Lactamases/sangue , beta-Lactamases/genética , Criança , Pré-Escolar , Croácia/epidemiologia , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Escherichia coli Uropatogênica/classificação
18.
Microb Drug Resist ; 17(2): 135-40, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21288137

RESUMO

Of forty-seven extended-spectrum cephalosporin-resistant Escherichia coli isolates, collected from children at the Children's Hospital in 2006 (Tunis, Tunisia), we analyzed 32 isolates that were genotypically different by enterobacterial repetitive intergenic consensus -polymerase chain reaction. For all isolates, the double-disk diffusion test revealed synergy between clavulanate and cefotaxime and/or ceftazidime, suggesting the production of extended-spectrum beta-lactamases. Polymerase chain reaction experiments, performed on plasmid DNA, and sequencing revealed the presence of bla(TEM-1B) (26 isolates, 81%), bla(TEM-34(IRT-6)) (3 isolates, 9%), bla(SHV-12) (2 isolates, 6%), and bla(CTX-M-15) (31 isolates, 97%). Further, the insertion sequence ISEcp1 was found upstream from the bla(CTX-M-15) gene in 11 isolates. The bla genes were found alone or in various combinations in a single isolate. bla(TEM-1B) and bla(CTX-M-15) genes were detected in 26 out of the 32 isolates. Three isolates harbored both bla(TEM-34(IRT-6)) and bla(CTX-M-15). bla(SHV-12) was identified either alone or with bla(CTX-M-15) in a single isolate. Our investigation showed the dominance of CTX-M-type extended-spectrum beta-lactamases, with CTX-M-15 particularly common, and to our best knowledge, this is the first report of the coexistence of CTX-M-15 and IRT-6 in E. coli isolates from children in Tunisia.


Assuntos
Antibacterianos/farmacologia , DNA Bacteriano/sangue , Infecções por Escherichia coli/genética , Escherichia coli/genética , Escherichia coli/isolamento & purificação , beta-Lactamases/genética , Técnicas de Tipagem Bacteriana , Cefotaxima/farmacologia , Ceftazidima/farmacologia , Criança , Ácido Clavulânico/farmacologia , Elementos de DNA Transponíveis/genética , DNA Intergênico/análise , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Farmacorresistência Bacteriana , Escherichia coli/classificação , Escherichia coli/efeitos dos fármacos , Escherichia coli/metabolismo , Infecções por Escherichia coli/tratamento farmacológico , Infecções por Escherichia coli/microbiologia , Fezes/microbiologia , Humanos , Plasmídeos/metabolismo , Reação em Cadeia da Polimerase , Tunísia , beta-Lactamases/sangue
19.
J Chromatogr Sci ; 48(8): 669-74, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20819297

RESUMO

For the determination of in vivo beta-lactamase activity, a high-performance liquid chromatographic (HPLC) method was established, and the pharmacokinetics of beta-lactamase after intravenous administration to the rats was analyzed using this standardized HPLC method. The plasma samples containing beta-lactamase were reacted with ampicillin (substrate) and further processed to make them fluorescent. The fluorescent compound of interest was separated using HPLC at room temperature using the excitation and the emission wavelengths of 410 nm and 475 nm, respectively. For the pharmacokinetic studies, 252 mU of beta-lactamase solution was administered to the rats through the tail vein injection (n = 6). The blood samples were withdrawn from the tail vein at different time points and analyzed by HPLC for beta-lactamase activity. For the HPLC method of beta-lactamase in plasma samples, the peak area showed a good correlation within the concentration ranges of 0.126-12.6 mU/mL (10-1000 ng/mL). The coefficients of variations were within 0.56-6.24, and the percentage recovery were within 102-107. After the intravenous injection, plasma concentration at the time zero (C(p0)) was 11.47 +/- 0.48 mU/mL, and no beta-lactamase was detected 24 h after the injection. The volume of distribution (V(d)) was 22 mL. An elimination half-life (t(1/2)) of 4.12 +/- 0.5 h and AUC of 79.4 +/- 12.9 mU.hr/mL were also calculated. The HPLC fluorimetric method was a very sensitive and reproducible method for the detection of beta-lactamase in plasma. The disposition of beta-lactamase after intravenous administration followed one-compartment and first-order kinetics.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Fluorometria/métodos , beta-Lactamases/sangue , Animais , Área Sob a Curva , Estabilidade de Medicamentos , Feminino , Modelos Lineares , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , beta-Lactamases/química , beta-Lactamases/farmacocinética
20.
J Craniomaxillofac Surg ; 36(8): 468-72, 2008 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18783957

RESUMO

OBJECTIVES: It is unclear to what extent pharmacokinetic data from bone outside the facial area can be transferred to the maxillofacial area. The aim of this study was to evaluate the penetration characteristics of piperacillin-tazobactam into human jaw and hip bone as a model for different bone characteristics. MATERIALS AND METHODS: The drug was administered at the start of surgery in a single 15-min intravenous infusion dose (4g piperacillin & 0.5g tazobactam i.v.). Plasma and bone samples of ten patients were analyzed. RESULTS: Mean concentration of piperacillin in plasma was 309microg/ml at 0.5h declining at 4h to 14microg/ml. The respective values for tazobactam were 34microg/ml declining to 2.8microg/ml. The piperacillin-tazobactam ratio dropped during the study interval from 0.5h: 9.2%+/-0.8 to 2h: 7.2%+/-1.1 and 4h: 4.9%+/-0.7. Mean bone concentrations of piperacillin and tazobactam were 9.0+/-11.6microg/g and 1.2+/-1.3microg/g, respectively. Mean penetration ratios for all bone samples were 15% (+/-17) for piperacillin and 13% (+/-14) for tazobactam without a difference between bone of different origin. DISCUSSION: Piperacillin-tazobactam levels in jaw bone tissue after a single dose are sufficient to assure antibacterial activity of the combination and are above the minimal inhibitory concentrations of the most relevant pathogens in head and neck surgery. Our data suggests the use of piperacillin-tazobactam as an alternative for the therapy of severe infections of the head and neck.


Assuntos
Antibacterianos/farmacocinética , Arcada Osseodentária/efeitos dos fármacos , Ossos Pélvicos/efeitos dos fármacos , beta-Lactamases/farmacocinética , Adolescente , Adulto , Antibacterianos/sangue , Combinação de Medicamentos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Ácido Penicilânico/análogos & derivados , Ácido Penicilânico/sangue , Ácido Penicilânico/farmacocinética , Piperacilina/sangue , Piperacilina/farmacocinética , Combinação Piperacilina e Tazobactam , Distribuição Tecidual , Adulto Jovem , Inibidores de beta-Lactamases , beta-Lactamases/sangue
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA