Avaliação de múltiplos mecanismos de resistência associados em isolados clínicos de Klebsiella pneumoniae resistente aos carbapenêmicos
Autor(a) principal: | |
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Data de Publicação: | 2015 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/6033 |
Resumo: | Antimicrobial resistance is considered a serious public health problem worldwide and complicates the treatment of infections caused by resistant microorganisms. The carbapenems are antimicrobial agents considered the last resource for treatment of severe infections caused by Klebsiella pneumoniae and the resistance to β-lactams can result in the accumulation of different resistance mechanisms (carbapenemases, efflux pump and loss of porins). This study aimed to evaluate multiple resistance mechanisms in 27 clinical isolates of K. pneumoniae resistant to carbapenems coming from the University Hospital of Santa Maria-RS from July 2013 to August 2014. These isolates were evaluated the susceptibility profiles through broth microdilution against ciprofloxacin, imipenem, ertapenem, meropenem, cefepime, ceftazidime and cefoxitin. Carbapenemase detection was performed through phenotypic tests with combined disc test with phenylboronic acid (AFB) and ethylenediaminetetraacetic acid (EDTA) and Blue-Carba test. In addition, genotypic tests to detect genes enconding carbapenemase were performed. Efflux pump was evaluated by broth microdilution together with efflux pump inhibitor and loss of porins were evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). High levels of resistance verified by the minimum inhibitory concentration (MIC) 50 and 90 for ciprofloxacin (64 and 128μg/mL), imipenem (32 to >128μg/mL), ertapenem (>128 and >128μg/mL), meropenem (128 and >128 μg/mL), cefepime (>128 and >128 μg/mL), ceftazidime (64 and 128μg/mL) and cefoxitin (128 and >128 μg/mL), respectively. In the resistance through carbapenemases production, 89% of the clinical isolates showed blaKPC gene and no clinical isolated showed genes encoding the metallo- β-lactamases. It was observed that the Blue-Carba test and combined disc test with AFB showed 100% concordance, while the combined disc test with EDTA showed high number of false positive (48%) when compared with genotypic test. Four isolates showed phenotypic profile consistent with the presence of efflux pump and all clinical isolates had lost one or both porins, being that in three isolated, this was the only resistance mechanism found. In 14% of the isolates can observe simultaneously observe the presence of three resistance mechanisms. Consequently, it is of fundamental scientific interest that studies are conducted in order to investigate and understand the mechanisms involved in resistance to carbapenems in order to assist strategies of prevention and infection control. |
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Avaliação de múltiplos mecanismos de resistência associados em isolados clínicos de Klebsiella pneumoniae resistente aos carbapenêmicosEvaluation of multiples associated resistance mechanisms in clinical isolates of K. pneumoniae resistant to carbapenemsCarbapenêmicosMecanismos de resistênciaKlebsiella pneumoniaePerda de porinasBomba de efluxoCarbapenemasesCarbapenensResistance mechanismKlebsiella pneumoniaeLoss of porinsEfflux pumpCarbapenemasesCNPQ::CIENCIAS DA SAUDE::FARMACIAAntimicrobial resistance is considered a serious public health problem worldwide and complicates the treatment of infections caused by resistant microorganisms. The carbapenems are antimicrobial agents considered the last resource for treatment of severe infections caused by Klebsiella pneumoniae and the resistance to β-lactams can result in the accumulation of different resistance mechanisms (carbapenemases, efflux pump and loss of porins). This study aimed to evaluate multiple resistance mechanisms in 27 clinical isolates of K. pneumoniae resistant to carbapenems coming from the University Hospital of Santa Maria-RS from July 2013 to August 2014. These isolates were evaluated the susceptibility profiles through broth microdilution against ciprofloxacin, imipenem, ertapenem, meropenem, cefepime, ceftazidime and cefoxitin. Carbapenemase detection was performed through phenotypic tests with combined disc test with phenylboronic acid (AFB) and ethylenediaminetetraacetic acid (EDTA) and Blue-Carba test. In addition, genotypic tests to detect genes enconding carbapenemase were performed. Efflux pump was evaluated by broth microdilution together with efflux pump inhibitor and loss of porins were evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). High levels of resistance verified by the minimum inhibitory concentration (MIC) 50 and 90 for ciprofloxacin (64 and 128μg/mL), imipenem (32 to >128μg/mL), ertapenem (>128 and >128μg/mL), meropenem (128 and >128 μg/mL), cefepime (>128 and >128 μg/mL), ceftazidime (64 and 128μg/mL) and cefoxitin (128 and >128 μg/mL), respectively. In the resistance through carbapenemases production, 89% of the clinical isolates showed blaKPC gene and no clinical isolated showed genes encoding the metallo- β-lactamases. It was observed that the Blue-Carba test and combined disc test with AFB showed 100% concordance, while the combined disc test with EDTA showed high number of false positive (48%) when compared with genotypic test. Four isolates showed phenotypic profile consistent with the presence of efflux pump and all clinical isolates had lost one or both porins, being that in three isolated, this was the only resistance mechanism found. In 14% of the isolates can observe simultaneously observe the presence of three resistance mechanisms. Consequently, it is of fundamental scientific interest that studies are conducted in order to investigate and understand the mechanisms involved in resistance to carbapenems in order to assist strategies of prevention and infection control.Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorA resistência aos antimicrobianos é considerada um grave problema de saúde pública em âmbito mundial e dificulta o tratamento de infecções causadas por microrganismos resistentes. Os carbapenêmicos são os antibacterianos considerados último recurso para o tratamento de infecções graves causadas por Klebsiella pneumoniae e a resistência a esse grupo de β-lactâmicos pode resultar da acumulação de diferentes mecanismos de resistência (carbapenemases, bomba de efluxo e perdas de porinas). Este trabalho teve como objetivo avaliar os múltiplos mecanismos de resistência de 27 isolados clínicos de K. pneumoniae resistente a carbapenêmicos oriundos do Hospital Universitário de Santa Maria-RS no período de julho de 2013 a agosto de 2014. Foram avaliados os perfis de suscetibilidade desses isolados através de microdiluição em caldo frente aos antimicrobianos ciprofloxacino, imipenem, ertapenem, meropenem, cefepima, ceftazidima e cefoxetina. A detecção de carbapenemases foi realizada através de testes fenotípicos com a utilização do disco de antimicrobiano associado com os inibidores ácido fenilborônico (AFB) e ácido etilenodiamino tetra-acético (EDTA) e através do teste Blue-Carba. Também foram realizados testes genotípicos para detectar os genes que codificam as carbapenemases. Bombas de efluxo foram avaliadas através de microdiluição em caldo juntamente com inibidor de bomba de efluxo e a perda de porinas foi avaliada pela eletroforese em gel de sulfato de dodecilo de sódio-poliacrilamida (SDS-PAGE). Altos níveis de resistência foram verificados nesses isolados pela concentração inibitória mínima (CIM) 50 e 90 para ciprofloxacino (64 e 128μg/mL), imipenem (32 e >128μg/mL), ertapenem (>128 e >128μg/mL), meropenem (128 e >128 μg/mL), cefepima (>128 e >128μg/mL), ceftazidima (64 e 128μg/mL) e cefoxitina (128 e >128 μg/mL), respectivamente. Na resistência através da produção de carbapenemases, 89% dos isolados apresentaram o gene blaKPC e nenhum isolado apresentou genes que codificam as metalo-β-lactamases. Foi observado que os testes Blue-Carba e disco combinado com AFB apresentaram 100% de concordância, enquanto o teste de disco combinado com EDTA apresentou elevado número de falso-positivos (48%) quando comparados com o teste genotípico. Quatro isolados apresentaram perfil fenotípico compatível com a presença de bomba de efluxo e todos os isolados apresentaram perda de uma ou ambas porinas, sendo que em três isolados, esse foi o único mecanismo de resistência encontrado. Em 14% dos isolados pode-se observar concomitantemente a presença dos três mecanismos de resistência. Devido ao exposto, é de fundamental interesse científico que estudos sejam realizados para investigar e compreender os mecanismos envolvidos na resistência aos carbapenêmicos, a fim de auxiliar em estratégias de prevenção e controle de infecção.Universidade Federal de Santa MariaBRFarmáciaUFSMPrograma de Pós-Graduação em Ciências FarmacêuticasCampos, Marli Matiko Anraku dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4737495P6Trindade, Priscila de Arrudahttp://lattes.cnpq.br/0124362929241308Matter, Leticia Beatrizhttp://lattes.cnpq.br/4288660833939683Loreto, érico Silva dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4713905Z8Dalmolin, Tanise Vendruscolo2016-03-312016-03-312015-08-21info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfDALMOLIN, Tanise Vendruscolo. EVALUATION OF MULTIPLES ASSOCIATED RESISTANCE MECHANISMS IN CLINICAL ISOLATES OF K. pneumoniae RESISTANT TO CARBAPENEMS. 2015. 60 f. Dissertação (Mestrado em Farmacologia) - Universidade Federal de Santa Maria, Santa Maria, 2015.http://repositorio.ufsm.br/handle/1/6033porinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2022-10-11T17:43:13Zoai:repositorio.ufsm.br:1/6033Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2022-10-11T17:43:13Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.none.fl_str_mv |
Avaliação de múltiplos mecanismos de resistência associados em isolados clínicos de Klebsiella pneumoniae resistente aos carbapenêmicos Evaluation of multiples associated resistance mechanisms in clinical isolates of K. pneumoniae resistant to carbapenems |
title |
Avaliação de múltiplos mecanismos de resistência associados em isolados clínicos de Klebsiella pneumoniae resistente aos carbapenêmicos |
spellingShingle |
Avaliação de múltiplos mecanismos de resistência associados em isolados clínicos de Klebsiella pneumoniae resistente aos carbapenêmicos Dalmolin, Tanise Vendruscolo Carbapenêmicos Mecanismos de resistência Klebsiella pneumoniae Perda de porinas Bomba de efluxo Carbapenemases Carbapenens Resistance mechanism Klebsiella pneumoniae Loss of porins Efflux pump Carbapenemases CNPQ::CIENCIAS DA SAUDE::FARMACIA |
title_short |
Avaliação de múltiplos mecanismos de resistência associados em isolados clínicos de Klebsiella pneumoniae resistente aos carbapenêmicos |
title_full |
Avaliação de múltiplos mecanismos de resistência associados em isolados clínicos de Klebsiella pneumoniae resistente aos carbapenêmicos |
title_fullStr |
Avaliação de múltiplos mecanismos de resistência associados em isolados clínicos de Klebsiella pneumoniae resistente aos carbapenêmicos |
title_full_unstemmed |
Avaliação de múltiplos mecanismos de resistência associados em isolados clínicos de Klebsiella pneumoniae resistente aos carbapenêmicos |
title_sort |
Avaliação de múltiplos mecanismos de resistência associados em isolados clínicos de Klebsiella pneumoniae resistente aos carbapenêmicos |
author |
Dalmolin, Tanise Vendruscolo |
author_facet |
Dalmolin, Tanise Vendruscolo |
author_role |
author |
dc.contributor.none.fl_str_mv |
Campos, Marli Matiko Anraku de http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4737495P6 Trindade, Priscila de Arruda http://lattes.cnpq.br/0124362929241308 Matter, Leticia Beatriz http://lattes.cnpq.br/4288660833939683 Loreto, érico Silva de http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4713905Z8 |
dc.contributor.author.fl_str_mv |
Dalmolin, Tanise Vendruscolo |
dc.subject.por.fl_str_mv |
Carbapenêmicos Mecanismos de resistência Klebsiella pneumoniae Perda de porinas Bomba de efluxo Carbapenemases Carbapenens Resistance mechanism Klebsiella pneumoniae Loss of porins Efflux pump Carbapenemases CNPQ::CIENCIAS DA SAUDE::FARMACIA |
topic |
Carbapenêmicos Mecanismos de resistência Klebsiella pneumoniae Perda de porinas Bomba de efluxo Carbapenemases Carbapenens Resistance mechanism Klebsiella pneumoniae Loss of porins Efflux pump Carbapenemases CNPQ::CIENCIAS DA SAUDE::FARMACIA |
description |
Antimicrobial resistance is considered a serious public health problem worldwide and complicates the treatment of infections caused by resistant microorganisms. The carbapenems are antimicrobial agents considered the last resource for treatment of severe infections caused by Klebsiella pneumoniae and the resistance to β-lactams can result in the accumulation of different resistance mechanisms (carbapenemases, efflux pump and loss of porins). This study aimed to evaluate multiple resistance mechanisms in 27 clinical isolates of K. pneumoniae resistant to carbapenems coming from the University Hospital of Santa Maria-RS from July 2013 to August 2014. These isolates were evaluated the susceptibility profiles through broth microdilution against ciprofloxacin, imipenem, ertapenem, meropenem, cefepime, ceftazidime and cefoxitin. Carbapenemase detection was performed through phenotypic tests with combined disc test with phenylboronic acid (AFB) and ethylenediaminetetraacetic acid (EDTA) and Blue-Carba test. In addition, genotypic tests to detect genes enconding carbapenemase were performed. Efflux pump was evaluated by broth microdilution together with efflux pump inhibitor and loss of porins were evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). High levels of resistance verified by the minimum inhibitory concentration (MIC) 50 and 90 for ciprofloxacin (64 and 128μg/mL), imipenem (32 to >128μg/mL), ertapenem (>128 and >128μg/mL), meropenem (128 and >128 μg/mL), cefepime (>128 and >128 μg/mL), ceftazidime (64 and 128μg/mL) and cefoxitin (128 and >128 μg/mL), respectively. In the resistance through carbapenemases production, 89% of the clinical isolates showed blaKPC gene and no clinical isolated showed genes encoding the metallo- β-lactamases. It was observed that the Blue-Carba test and combined disc test with AFB showed 100% concordance, while the combined disc test with EDTA showed high number of false positive (48%) when compared with genotypic test. Four isolates showed phenotypic profile consistent with the presence of efflux pump and all clinical isolates had lost one or both porins, being that in three isolated, this was the only resistance mechanism found. In 14% of the isolates can observe simultaneously observe the presence of three resistance mechanisms. Consequently, it is of fundamental scientific interest that studies are conducted in order to investigate and understand the mechanisms involved in resistance to carbapenems in order to assist strategies of prevention and infection control. |
publishDate |
2015 |
dc.date.none.fl_str_mv |
2015-08-21 2016-03-31 2016-03-31 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
DALMOLIN, Tanise Vendruscolo. EVALUATION OF MULTIPLES ASSOCIATED RESISTANCE MECHANISMS IN CLINICAL ISOLATES OF K. pneumoniae RESISTANT TO CARBAPENEMS. 2015. 60 f. Dissertação (Mestrado em Farmacologia) - Universidade Federal de Santa Maria, Santa Maria, 2015. http://repositorio.ufsm.br/handle/1/6033 |
identifier_str_mv |
DALMOLIN, Tanise Vendruscolo. EVALUATION OF MULTIPLES ASSOCIATED RESISTANCE MECHANISMS IN CLINICAL ISOLATES OF K. pneumoniae RESISTANT TO CARBAPENEMS. 2015. 60 f. Dissertação (Mestrado em Farmacologia) - Universidade Federal de Santa Maria, Santa Maria, 2015. |
url |
http://repositorio.ufsm.br/handle/1/6033 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
dc.format.none.fl_str_mv |
application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria BR Farmácia UFSM Programa de Pós-Graduação em Ciências Farmacêuticas |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria BR Farmácia UFSM Programa de Pós-Graduação em Ciências Farmacêuticas |
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reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
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Universidade Federal de Santa Maria (UFSM) |
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UFSM |
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UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
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Manancial - Repositório Digital da UFSM |
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Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
atendimento.sib@ufsm.br||tedebc@gmail.com |
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