Caracterização fenotípica e molecular dos mecanismos de resistência aos beta-lactâmicos em linhagens clínicas de Klebsiella pneumoniae isoladas do Amazonas
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/11839 |
Resumo: | Klebsiella pneumoniae resistant to beta-lactams are present in hospitals worldwide and reduce treatment alternatives. In this study, 40 K. pneumoniae isolates were characterized. All of them were resistant to beta-lactam antibiotics and classified as multidrug resistant. Among these, eight isolates were resistant to carbapenems and six of which had the gene blaKPC-2, encoding the enzyme Klebsiella pneumoniae carbapenemase (KPC). The blaTEM and blaCTX-M genes were found in 80% of the samples. The 40 isolates were grouped into 12 pulses (A to L) according to the genetic similarity determined by the DNA macro-restriction followed by pulsed-field gel electrophoresis. A total of 20 isolates were classified as pulsotype A, which was subdivided into 13 subtypes (A1 to A13), with subtype A1 being the most prevalent. Representatives of pulsotype A belong to the ST11, clonal complex CC258. In this study, we identified four new STs: ST2540, ST2308, ST2258, and ST2307. AMKP5 and AMKP2, belonging to subtypes A7 and A8, respectively, are resistant to carbapenems but did not present the blaKPC gene. Sequencing of the AMKP5 genome indicated mutations in the C-terminal region of the OmpK36 porin which may alter the entry of carbapenems into the cell. Genome sequencing of the AMKP10 isolate identified the plasmid pAMKP10, with 48.454 bp belonging to the IncX5 incompatibility group. In this plasmid, the blaKPC-2 gene was found in a non-Tn4401 genetic element (NTEKPC) containing tnpA-ISEc63-like / tnpR-Tn3 / tnpA-ISKpn27 / blaKPC-2 / ΔISKpn6, based on Tn1722. All isolates with the blaKPC-2 gene had the same plasmid. Conjugation of this plasmid to Escherichia coli J53 occurred from ST11 isolates with low conjugation frequencies (8.23 x 10-12 to 1.31 x 10-7) resulting in resistance to carbapenems. Finally, the outbreak of carbapenem-resistant K. pneumoniae resistant of this Amazonian hospital involved the transfer of the NTEKPC-containing pAMKP10 between two lineages (ST11 and ST2302) but was largely due to the clonal dissemination of the ST11 endemic lineage. |
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Souza, Rosineide Cardoso deCamargo, Ilana Lopes Baratella da Cunhahttp://lattes.cnpq.br/4104096171600845http://lattes.cnpq.br/5739753892325187ff125bc7-7846-4cdf-8c1b-c73b9a8413e92019-09-13T19:50:32Z2019-09-13T19:50:32Z2018-10-25SOUZA, Rosineide Cardoso de. Caracterização fenotípica e molecular dos mecanismos de resistência aos beta-lactâmicos em linhagens clínicas de Klebsiella pneumoniae isoladas do Amazonas. 2018. Tese (Doutorado em Genética Evolutiva e Biologia Molecular) – Universidade Federal de São Carlos, São Carlos, 2018. Disponível em: https://repositorio.ufscar.br/handle/ufscar/11839.https://repositorio.ufscar.br/handle/ufscar/11839Klebsiella pneumoniae resistant to beta-lactams are present in hospitals worldwide and reduce treatment alternatives. In this study, 40 K. pneumoniae isolates were characterized. All of them were resistant to beta-lactam antibiotics and classified as multidrug resistant. Among these, eight isolates were resistant to carbapenems and six of which had the gene blaKPC-2, encoding the enzyme Klebsiella pneumoniae carbapenemase (KPC). The blaTEM and blaCTX-M genes were found in 80% of the samples. The 40 isolates were grouped into 12 pulses (A to L) according to the genetic similarity determined by the DNA macro-restriction followed by pulsed-field gel electrophoresis. A total of 20 isolates were classified as pulsotype A, which was subdivided into 13 subtypes (A1 to A13), with subtype A1 being the most prevalent. Representatives of pulsotype A belong to the ST11, clonal complex CC258. In this study, we identified four new STs: ST2540, ST2308, ST2258, and ST2307. AMKP5 and AMKP2, belonging to subtypes A7 and A8, respectively, are resistant to carbapenems but did not present the blaKPC gene. Sequencing of the AMKP5 genome indicated mutations in the C-terminal region of the OmpK36 porin which may alter the entry of carbapenems into the cell. Genome sequencing of the AMKP10 isolate identified the plasmid pAMKP10, with 48.454 bp belonging to the IncX5 incompatibility group. In this plasmid, the blaKPC-2 gene was found in a non-Tn4401 genetic element (NTEKPC) containing tnpA-ISEc63-like / tnpR-Tn3 / tnpA-ISKpn27 / blaKPC-2 / ΔISKpn6, based on Tn1722. All isolates with the blaKPC-2 gene had the same plasmid. Conjugation of this plasmid to Escherichia coli J53 occurred from ST11 isolates with low conjugation frequencies (8.23 x 10-12 to 1.31 x 10-7) resulting in resistance to carbapenems. Finally, the outbreak of carbapenem-resistant K. pneumoniae resistant of this Amazonian hospital involved the transfer of the NTEKPC-containing pAMKP10 between two lineages (ST11 and ST2302) but was largely due to the clonal dissemination of the ST11 endemic lineage.Klebsiella pneumoniae resistentes aos beta-lactâmicos estão presentes em hospitais do mundo todo e reduzem as alternativas de tratamento. Nesse estudo, 40 isolados de K. pneumoniae foram caracterizados. Todos foram classificados como multirresistentes e com resistência aos antibióticos beta-lactâmicos. Dentre estes, oito amostras são resistentes aos carbapenêmicos e seis delas têm o gene blaKPC-2, que codifica a enzima Klebsiella pneumoniae carbapenemase (KPC). Os genes blaTEM e blaCTX-M foram encontrados em 80% das amostras. Os 40 isolados foram agrupados em 12 pulsotipos (A a L) de acordo com a similaridade genética determinada pela macrorrestrição do DNA seguida de eletroforese em gel de campos pulsados. Um total de 20 isolados foram classificados como pulsotipo A, o qual foi subdividido em 13 subtipos (A1 a A13), sendo o subtipo A1 o prevalente. Representantes do pulsotipo A pertencem ao ST11, complexo clonal CC258. Neste estudo, quatro novos STs foram identificados: ST2540, ST2308, ST2258 e ST2307. AMKP5 e AMKP2, dos subtipos A7 e A8, respectivamente, são resistentes aos carbapenêmicos, mas não apresentaram o gene blaKPC. O sequenciamento do genoma da AMKP5 indicou mutações na região C-terminal da porina OmpK36 que pode alterar a entrada dos carbapenêmicos na célula. O sequenciamento do genoma da amostra AMKP10 identificou o plasmídeo pAMKP10, com 48,454 pb pertencente ao grupo de incompatibilidade IncX5. Neste plasmídeo, o gene blaKPC foi encontrado em um ambiente genético diferente do Tn4401 (NTEKPC do inglês, non-Tn4401 genetic element) contendo tnpA-ISEc63-like/tnpR-Tn3/tnpA-ISKpn27/blaKPC-2/ΔISKpn6, baseado no Tn1722. Todos os isolados com o gene blaKPC-2 apresentaram o mesmo plasmídeo. A conjugação deste plasmídeo para Escherichia coli J53 ocorreu a partir dos isolados ST11 com baixas frequências de conjugação (de 8,23 x 10-12 a 1,31 x 10-7) resultando em resistência aos carbapenêmicos. Por fim, observou-se que o surto de K. pneumoniae resistentes aos carbapenêmicos deste hospital Amazonense envolveu a transferência do pAMKP10 contendo NTEKPC entre duas linhagens (ST11 e ST2302), mas ocorreu em grande parte devido à disseminação clonal da linhagem endêmica ST11.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundação de Amparo à Pesquisa do Estado do Amazonas (FAPEAM)FAPEAM: 24868.484.14650.07112014-35526FAPESP: 13/07600-l3CNPq: 131314/2018-1porUniversidade Federal de São CarlosCâmpus São CarlosPrograma de Pós-Graduação em Genética Evolutiva e Biologia Molecular - PPGGEvUFSCarKlebsiella pneumoniaeblaKPC-2NTEKPCIncX5CarbapenemCIENCIAS BIOLOGICAS::BIOQUIMICACIENCIAS BIOLOGICAS::GENETICACIENCIAS BIOLOGICAS::MICROBIOLOGIACaracterização fenotípica e molecular dos mecanismos de resistência aos beta-lactâmicos em linhagens clínicas de Klebsiella pneumoniae isoladas do AmazonasPhenotypic and molecular characterization of beta-lactam resistance mechanisms in clinical strains of Klebsiella pneumoniae isolated from Amazonasinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis6 meses após a data da defesa22028037-951f-464b-bc2d-bce3c9e94ea4info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALRosineide Cardoso de Souza.pdfRosineide Cardoso de Souza.pdfapplication/pdf1974442https://repositorio.ufscar.br/bitstream/ufscar/11839/1/Rosineide%20Cardoso%20de%20Souza.pdfa0c9abc2cb3da25daf526e606d7495d8MD51carta-comprovante 2.pdfcarta-comprovante 2.pdfCarta comprovante de deposito da versão final da teseapplication/pdf89372https://repositorio.ufscar.br/bitstream/ufscar/11839/3/carta-comprovante%202.pdf33c79ecc26d904fcfd130c4bbba8733cMD53LICENSElicense.txtlicense.txttext/plain; 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dc.title.por.fl_str_mv |
Caracterização fenotípica e molecular dos mecanismos de resistência aos beta-lactâmicos em linhagens clínicas de Klebsiella pneumoniae isoladas do Amazonas |
dc.title.alternative.eng.fl_str_mv |
Phenotypic and molecular characterization of beta-lactam resistance mechanisms in clinical strains of Klebsiella pneumoniae isolated from Amazonas |
title |
Caracterização fenotípica e molecular dos mecanismos de resistência aos beta-lactâmicos em linhagens clínicas de Klebsiella pneumoniae isoladas do Amazonas |
spellingShingle |
Caracterização fenotípica e molecular dos mecanismos de resistência aos beta-lactâmicos em linhagens clínicas de Klebsiella pneumoniae isoladas do Amazonas Souza, Rosineide Cardoso de Klebsiella pneumoniae blaKPC-2 NTEKPC IncX5 Carbapenem CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS BIOLOGICAS::GENETICA CIENCIAS BIOLOGICAS::MICROBIOLOGIA |
title_short |
Caracterização fenotípica e molecular dos mecanismos de resistência aos beta-lactâmicos em linhagens clínicas de Klebsiella pneumoniae isoladas do Amazonas |
title_full |
Caracterização fenotípica e molecular dos mecanismos de resistência aos beta-lactâmicos em linhagens clínicas de Klebsiella pneumoniae isoladas do Amazonas |
title_fullStr |
Caracterização fenotípica e molecular dos mecanismos de resistência aos beta-lactâmicos em linhagens clínicas de Klebsiella pneumoniae isoladas do Amazonas |
title_full_unstemmed |
Caracterização fenotípica e molecular dos mecanismos de resistência aos beta-lactâmicos em linhagens clínicas de Klebsiella pneumoniae isoladas do Amazonas |
title_sort |
Caracterização fenotípica e molecular dos mecanismos de resistência aos beta-lactâmicos em linhagens clínicas de Klebsiella pneumoniae isoladas do Amazonas |
author |
Souza, Rosineide Cardoso de |
author_facet |
Souza, Rosineide Cardoso de |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/5739753892325187 |
dc.contributor.author.fl_str_mv |
Souza, Rosineide Cardoso de |
dc.contributor.advisor1.fl_str_mv |
Camargo, Ilana Lopes Baratella da Cunha |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/4104096171600845 |
dc.contributor.authorID.fl_str_mv |
ff125bc7-7846-4cdf-8c1b-c73b9a8413e9 |
contributor_str_mv |
Camargo, Ilana Lopes Baratella da Cunha |
dc.subject.eng.fl_str_mv |
Klebsiella pneumoniae blaKPC-2 NTEKPC IncX5 Carbapenem |
topic |
Klebsiella pneumoniae blaKPC-2 NTEKPC IncX5 Carbapenem CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS BIOLOGICAS::GENETICA CIENCIAS BIOLOGICAS::MICROBIOLOGIA |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS BIOLOGICAS::GENETICA CIENCIAS BIOLOGICAS::MICROBIOLOGIA |
description |
Klebsiella pneumoniae resistant to beta-lactams are present in hospitals worldwide and reduce treatment alternatives. In this study, 40 K. pneumoniae isolates were characterized. All of them were resistant to beta-lactam antibiotics and classified as multidrug resistant. Among these, eight isolates were resistant to carbapenems and six of which had the gene blaKPC-2, encoding the enzyme Klebsiella pneumoniae carbapenemase (KPC). The blaTEM and blaCTX-M genes were found in 80% of the samples. The 40 isolates were grouped into 12 pulses (A to L) according to the genetic similarity determined by the DNA macro-restriction followed by pulsed-field gel electrophoresis. A total of 20 isolates were classified as pulsotype A, which was subdivided into 13 subtypes (A1 to A13), with subtype A1 being the most prevalent. Representatives of pulsotype A belong to the ST11, clonal complex CC258. In this study, we identified four new STs: ST2540, ST2308, ST2258, and ST2307. AMKP5 and AMKP2, belonging to subtypes A7 and A8, respectively, are resistant to carbapenems but did not present the blaKPC gene. Sequencing of the AMKP5 genome indicated mutations in the C-terminal region of the OmpK36 porin which may alter the entry of carbapenems into the cell. Genome sequencing of the AMKP10 isolate identified the plasmid pAMKP10, with 48.454 bp belonging to the IncX5 incompatibility group. In this plasmid, the blaKPC-2 gene was found in a non-Tn4401 genetic element (NTEKPC) containing tnpA-ISEc63-like / tnpR-Tn3 / tnpA-ISKpn27 / blaKPC-2 / ΔISKpn6, based on Tn1722. All isolates with the blaKPC-2 gene had the same plasmid. Conjugation of this plasmid to Escherichia coli J53 occurred from ST11 isolates with low conjugation frequencies (8.23 x 10-12 to 1.31 x 10-7) resulting in resistance to carbapenems. Finally, the outbreak of carbapenem-resistant K. pneumoniae resistant of this Amazonian hospital involved the transfer of the NTEKPC-containing pAMKP10 between two lineages (ST11 and ST2302) but was largely due to the clonal dissemination of the ST11 endemic lineage. |
publishDate |
2018 |
dc.date.issued.fl_str_mv |
2018-10-25 |
dc.date.accessioned.fl_str_mv |
2019-09-13T19:50:32Z |
dc.date.available.fl_str_mv |
2019-09-13T19:50:32Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
SOUZA, Rosineide Cardoso de. Caracterização fenotípica e molecular dos mecanismos de resistência aos beta-lactâmicos em linhagens clínicas de Klebsiella pneumoniae isoladas do Amazonas. 2018. Tese (Doutorado em Genética Evolutiva e Biologia Molecular) – Universidade Federal de São Carlos, São Carlos, 2018. Disponível em: https://repositorio.ufscar.br/handle/ufscar/11839. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/11839 |
identifier_str_mv |
SOUZA, Rosineide Cardoso de. Caracterização fenotípica e molecular dos mecanismos de resistência aos beta-lactâmicos em linhagens clínicas de Klebsiella pneumoniae isoladas do Amazonas. 2018. Tese (Doutorado em Genética Evolutiva e Biologia Molecular) – Universidade Federal de São Carlos, São Carlos, 2018. Disponível em: https://repositorio.ufscar.br/handle/ufscar/11839. |
url |
https://repositorio.ufscar.br/handle/ufscar/11839 |
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por |
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por |
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dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de São Carlos Câmpus São Carlos |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Genética Evolutiva e Biologia Molecular - PPGGEv |
dc.publisher.initials.fl_str_mv |
UFSCar |
publisher.none.fl_str_mv |
Universidade Federal de São Carlos Câmpus São Carlos |
dc.source.none.fl_str_mv |
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Universidade Federal de São Carlos (UFSCAR) |
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institution |
UFSCAR |
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collection |
Repositório Institucional da UFSCAR |
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Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR) |
repository.mail.fl_str_mv |
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1802136363841093632 |