Novel cassette assay to quantify the outer membrane permeability of five β-lactams simultaneously in carbapenem-resistant Klebsiella pneumoniae and Enterobactercloacae
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UFRGS |
Texto Completo: | http://hdl.handle.net/10183/229311 |
Resumo: | Poor penetration through the outer membrane (OM) of Gram-negative bacteria is a major barrier of antibiotic development. While β-lactam antibiotics are commonly used against Klebsiella pneumoniae and Enterobacter cloacae, there are limited data on OM permeability especially in K. pneumoniae. Here, we developed a novel cassette assay, which can simultaneously quantify the OM permeability to five β-lactams in carbapenem-resistant K. pneumoniae and E. cloacae. Both clinical isolates harbored a blaKPC-2 and several other β-lactamases. The OM permeability of each antibiotic was studied separately (“discrete assay”) and simultaneously (“cassette assay”) by determining the degradation of extracellular β-lactam concentrations via multiplex liquid chromatography-tandem mass spectrometry analyses. Our K. pneumoniae isolate was polymyxin resistant, whereas the E. cloacae was polymyxin susceptible. Imipenem penetrated the OM at least 7-fold faster than meropenem for both isolates. Imipenem penetrated E. cloacae at least 258-fold faster and K. pneumoniae 150-fold faster compared to aztreonam, cefepime, and ceftazidime. For our β-lactams, OM permeability was substantially higher in the E. cloacae compared to the K. pneumoniae isolate (except for aztreonam). This correlated with a higher OmpC porin production in E. cloacae, as determined by proteomics. The cassette and discrete assays showed comparable results, suggesting limited or no competition during influx through OM porins. This cassette assay allowed us, for the first time, to efficiently quantify the OM permeability of multiple β-lactams in carbapenem-resistant K. pneumoniae and E. cloacae. Characterizing the OM permeability presents a critical contribution to combating the antimicrobial resistance crisis and enables us to rationally optimize the use of β-lactam antibiotics. |
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Tae Hwan, KimZavascki, Alexandre PrehnBarth, Afonso LuisBülitta, Jürgen Bernd B.2021-09-01T04:24:34Z20202161-2129http://hdl.handle.net/10183/229311001130284Poor penetration through the outer membrane (OM) of Gram-negative bacteria is a major barrier of antibiotic development. While β-lactam antibiotics are commonly used against Klebsiella pneumoniae and Enterobacter cloacae, there are limited data on OM permeability especially in K. pneumoniae. Here, we developed a novel cassette assay, which can simultaneously quantify the OM permeability to five β-lactams in carbapenem-resistant K. pneumoniae and E. cloacae. Both clinical isolates harbored a blaKPC-2 and several other β-lactamases. The OM permeability of each antibiotic was studied separately (“discrete assay”) and simultaneously (“cassette assay”) by determining the degradation of extracellular β-lactam concentrations via multiplex liquid chromatography-tandem mass spectrometry analyses. Our K. pneumoniae isolate was polymyxin resistant, whereas the E. cloacae was polymyxin susceptible. Imipenem penetrated the OM at least 7-fold faster than meropenem for both isolates. Imipenem penetrated E. cloacae at least 258-fold faster and K. pneumoniae 150-fold faster compared to aztreonam, cefepime, and ceftazidime. For our β-lactams, OM permeability was substantially higher in the E. cloacae compared to the K. pneumoniae isolate (except for aztreonam). This correlated with a higher OmpC porin production in E. cloacae, as determined by proteomics. The cassette and discrete assays showed comparable results, suggesting limited or no competition during influx through OM porins. This cassette assay allowed us, for the first time, to efficiently quantify the OM permeability of multiple β-lactams in carbapenem-resistant K. pneumoniae and E. cloacae. Characterizing the OM permeability presents a critical contribution to combating the antimicrobial resistance crisis and enables us to rationally optimize the use of β-lactam antibiotics.application/pdfengmBio. Washington. Vol. 11, no. 1 (Jan./Fab. 2020), e03189-19, p. 1-15.Enterobacter cloacaeInfecções por enterobacteriaceaeKlebsiella pneumoniaeInfecções por KlebsiellaPermeabilidade da membrana celularEnterobacteriáceas resistentes a carbapenêmicosBeta-LactamasEnterobacter cloacaeKlebsiella pneumoniaeLC-MS/MSBeta-lactamsCarbapenem resistanceCarbapenemsCassette assayCephalosporinsMonobactamsOuter membranePermeabilityPolymyxin resistanceNovel cassette assay to quantify the outer membrane permeability of five β-lactams simultaneously in carbapenem-resistant Klebsiella pneumoniae and EnterobactercloacaeEstrangeiroinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFRGSinstname:Universidade Federal do Rio Grande do Sul (UFRGS)instacron:UFRGSTEXT001130284.pdf.txt001130284.pdf.txtExtracted Texttext/plain70735http://www.lume.ufrgs.br/bitstream/10183/229311/2/001130284.pdf.txt365a413b2b25327968b32c2b36de31d7MD52ORIGINAL001130284.pdfTexto completo (inglês)application/pdf3648363http://www.lume.ufrgs.br/bitstream/10183/229311/1/001130284.pdf5951fd7599c81a12ecbc3f787bf970d2MD5110183/2293112021-09-19 04:28:03.480743oai:www.lume.ufrgs.br:10183/229311Repositório de PublicaçõesPUBhttps://lume.ufrgs.br/oai/requestopendoar:2021-09-19T07:28:03Repositório Institucional da UFRGS - Universidade Federal do Rio Grande do Sul (UFRGS)false |
dc.title.pt_BR.fl_str_mv |
Novel cassette assay to quantify the outer membrane permeability of five β-lactams simultaneously in carbapenem-resistant Klebsiella pneumoniae and Enterobactercloacae |
title |
Novel cassette assay to quantify the outer membrane permeability of five β-lactams simultaneously in carbapenem-resistant Klebsiella pneumoniae and Enterobactercloacae |
spellingShingle |
Novel cassette assay to quantify the outer membrane permeability of five β-lactams simultaneously in carbapenem-resistant Klebsiella pneumoniae and Enterobactercloacae Tae Hwan, Kim Enterobacter cloacae Infecções por enterobacteriaceae Klebsiella pneumoniae Infecções por Klebsiella Permeabilidade da membrana celular Enterobacteriáceas resistentes a carbapenêmicos Beta-Lactamas Enterobacter cloacae Klebsiella pneumoniae LC-MS/MS Beta-lactams Carbapenem resistance Carbapenems Cassette assay Cephalosporins Monobactams Outer membrane Permeability Polymyxin resistance |
title_short |
Novel cassette assay to quantify the outer membrane permeability of five β-lactams simultaneously in carbapenem-resistant Klebsiella pneumoniae and Enterobactercloacae |
title_full |
Novel cassette assay to quantify the outer membrane permeability of five β-lactams simultaneously in carbapenem-resistant Klebsiella pneumoniae and Enterobactercloacae |
title_fullStr |
Novel cassette assay to quantify the outer membrane permeability of five β-lactams simultaneously in carbapenem-resistant Klebsiella pneumoniae and Enterobactercloacae |
title_full_unstemmed |
Novel cassette assay to quantify the outer membrane permeability of five β-lactams simultaneously in carbapenem-resistant Klebsiella pneumoniae and Enterobactercloacae |
title_sort |
Novel cassette assay to quantify the outer membrane permeability of five β-lactams simultaneously in carbapenem-resistant Klebsiella pneumoniae and Enterobactercloacae |
author |
Tae Hwan, Kim |
author_facet |
Tae Hwan, Kim Zavascki, Alexandre Prehn Barth, Afonso Luis Bülitta, Jürgen Bernd B. |
author_role |
author |
author2 |
Zavascki, Alexandre Prehn Barth, Afonso Luis Bülitta, Jürgen Bernd B. |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Tae Hwan, Kim Zavascki, Alexandre Prehn Barth, Afonso Luis Bülitta, Jürgen Bernd B. |
dc.subject.por.fl_str_mv |
Enterobacter cloacae Infecções por enterobacteriaceae Klebsiella pneumoniae Infecções por Klebsiella Permeabilidade da membrana celular Enterobacteriáceas resistentes a carbapenêmicos Beta-Lactamas |
topic |
Enterobacter cloacae Infecções por enterobacteriaceae Klebsiella pneumoniae Infecções por Klebsiella Permeabilidade da membrana celular Enterobacteriáceas resistentes a carbapenêmicos Beta-Lactamas Enterobacter cloacae Klebsiella pneumoniae LC-MS/MS Beta-lactams Carbapenem resistance Carbapenems Cassette assay Cephalosporins Monobactams Outer membrane Permeability Polymyxin resistance |
dc.subject.eng.fl_str_mv |
Enterobacter cloacae Klebsiella pneumoniae LC-MS/MS Beta-lactams Carbapenem resistance Carbapenems Cassette assay Cephalosporins Monobactams Outer membrane Permeability Polymyxin resistance |
description |
Poor penetration through the outer membrane (OM) of Gram-negative bacteria is a major barrier of antibiotic development. While β-lactam antibiotics are commonly used against Klebsiella pneumoniae and Enterobacter cloacae, there are limited data on OM permeability especially in K. pneumoniae. Here, we developed a novel cassette assay, which can simultaneously quantify the OM permeability to five β-lactams in carbapenem-resistant K. pneumoniae and E. cloacae. Both clinical isolates harbored a blaKPC-2 and several other β-lactamases. The OM permeability of each antibiotic was studied separately (“discrete assay”) and simultaneously (“cassette assay”) by determining the degradation of extracellular β-lactam concentrations via multiplex liquid chromatography-tandem mass spectrometry analyses. Our K. pneumoniae isolate was polymyxin resistant, whereas the E. cloacae was polymyxin susceptible. Imipenem penetrated the OM at least 7-fold faster than meropenem for both isolates. Imipenem penetrated E. cloacae at least 258-fold faster and K. pneumoniae 150-fold faster compared to aztreonam, cefepime, and ceftazidime. For our β-lactams, OM permeability was substantially higher in the E. cloacae compared to the K. pneumoniae isolate (except for aztreonam). This correlated with a higher OmpC porin production in E. cloacae, as determined by proteomics. The cassette and discrete assays showed comparable results, suggesting limited or no competition during influx through OM porins. This cassette assay allowed us, for the first time, to efficiently quantify the OM permeability of multiple β-lactams in carbapenem-resistant K. pneumoniae and E. cloacae. Characterizing the OM permeability presents a critical contribution to combating the antimicrobial resistance crisis and enables us to rationally optimize the use of β-lactam antibiotics. |
publishDate |
2020 |
dc.date.issued.fl_str_mv |
2020 |
dc.date.accessioned.fl_str_mv |
2021-09-01T04:24:34Z |
dc.type.driver.fl_str_mv |
Estrangeiro info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10183/229311 |
dc.identifier.issn.pt_BR.fl_str_mv |
2161-2129 |
dc.identifier.nrb.pt_BR.fl_str_mv |
001130284 |
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http://hdl.handle.net/10183/229311 |
dc.language.iso.fl_str_mv |
eng |
language |
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dc.relation.ispartof.pt_BR.fl_str_mv |
mBio. Washington. Vol. 11, no. 1 (Jan./Fab. 2020), e03189-19, p. 1-15. |
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info:eu-repo/semantics/openAccess |
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