Caracterização fenotípica e molecular da resistência a clindamicina em isolados clínicos de Staphylococcus aureus de um hospital terciário

Detalhes bibliográficos
Autor(a) principal: Bottega, Angelita
Data de Publicação: 2016
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações do UFSM
Texto Completo: http://repositorio.ufsm.br/handle/1/18165
Resumo: Staphylococcus aureus demonstrated over the time, the ability to develop resistance to the most antimicrobials. The mechanism of resistance to macrolides in S. aureus also reaches lincosamides and streptogramins B, featuring the designate MLSB resistance, whose expression can be constitutive (cMLSB) or inducible (iMLSB) and it is encoded mainly by the presence of erm genes (ermA, ermB and ermC) and msrA. The cMLSB resistance is easily detected by susceptibility testing used in routine laboratory, but iMLSB resistance identification is only possible by using an inducing agent. Thus, clindamycin therapy in cases of infection by isolated with iMLSB resistance can fail. Thus, this study aimed to characterize the phenotypic profile (cMLSB e iMLSB) and molecular (presence of ermA, ermB, ermC and mrsA genes) of 140 clinical isolates of methicillin-sensitive S. aureus (MSSA) and methicillin-resistant (MRSA), from patients admitted at University Hospital of Santa Maria (HUSM) in the period from April to December 2011.The inclusion criteria were all isolates identified as S. aureus by manual phenotypic methods (Gram stain, catalase test, coagulase, etc.) and automated (MicroScan® - Siemens), and excluded of the study repeated samples from the same patient. The MLSB phenotype was detected by approaching disk induction test (D-test), and from 140 strains, 25 had cMLSB phenotype and 11 the iMLSB phenotype. Of constitutive isolates, 20 were MRSA and 5 MSSA, while the inducible isolates 8 (5.8%) were MSSA and 3 (2.1%) MRSA. From these results, we performed the molecular characterization by Reaction technique Polymerase Chain (PCR) of all isolates that expressed the MLSB resistance in phenotypic test. Among these, ermA and ermC genes were prevalent, identified at 11 and 15 of the isolates, respectively. The gene ermB amplified in 4of 36 strains, while msrA in 13, and of these, 8 had an association with the erm genes surveyed. Despite the iMLSB phenotype have been less frequent than cMLSB, the realization of the D-test is important to detect these phenotypes, it may guide therapeutic procedures, helping to improve in the health of the patient, reducing the length of hospitalization and thus the burden on the health system. Moreover, the test is inexpensive and easy to run compared to the PCR technique, which despite its high sensitivity and specificity is not used in routine of all clinical laboratories due to high costs.
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spelling 2019-09-06T21:34:10Z2019-09-06T21:34:10Z2016-08-11http://repositorio.ufsm.br/handle/1/18165Staphylococcus aureus demonstrated over the time, the ability to develop resistance to the most antimicrobials. The mechanism of resistance to macrolides in S. aureus also reaches lincosamides and streptogramins B, featuring the designate MLSB resistance, whose expression can be constitutive (cMLSB) or inducible (iMLSB) and it is encoded mainly by the presence of erm genes (ermA, ermB and ermC) and msrA. The cMLSB resistance is easily detected by susceptibility testing used in routine laboratory, but iMLSB resistance identification is only possible by using an inducing agent. Thus, clindamycin therapy in cases of infection by isolated with iMLSB resistance can fail. Thus, this study aimed to characterize the phenotypic profile (cMLSB e iMLSB) and molecular (presence of ermA, ermB, ermC and mrsA genes) of 140 clinical isolates of methicillin-sensitive S. aureus (MSSA) and methicillin-resistant (MRSA), from patients admitted at University Hospital of Santa Maria (HUSM) in the period from April to December 2011.The inclusion criteria were all isolates identified as S. aureus by manual phenotypic methods (Gram stain, catalase test, coagulase, etc.) and automated (MicroScan® - Siemens), and excluded of the study repeated samples from the same patient. The MLSB phenotype was detected by approaching disk induction test (D-test), and from 140 strains, 25 had cMLSB phenotype and 11 the iMLSB phenotype. Of constitutive isolates, 20 were MRSA and 5 MSSA, while the inducible isolates 8 (5.8%) were MSSA and 3 (2.1%) MRSA. From these results, we performed the molecular characterization by Reaction technique Polymerase Chain (PCR) of all isolates that expressed the MLSB resistance in phenotypic test. Among these, ermA and ermC genes were prevalent, identified at 11 and 15 of the isolates, respectively. The gene ermB amplified in 4of 36 strains, while msrA in 13, and of these, 8 had an association with the erm genes surveyed. Despite the iMLSB phenotype have been less frequent than cMLSB, the realization of the D-test is important to detect these phenotypes, it may guide therapeutic procedures, helping to improve in the health of the patient, reducing the length of hospitalization and thus the burden on the health system. Moreover, the test is inexpensive and easy to run compared to the PCR technique, which despite its high sensitivity and specificity is not used in routine of all clinical laboratories due to high costs.Os Staphylococcus aureus demonstraram ao longo do tempo, capacidade de desenvolver resistência a maioria dos antimicrobianos. O mecanismo de resistência aos macrolídeos em S. aureus atinge também as lincosamidas e as estreptograminas B, caracterizando a denominada resistência MLSB, cuja expressão pode ser constitutiva (cMLSB) ou induzível (iMLSB) e é codificada principalmente pela presença dos genes erm (ermA, ermB e ermC) e msrA. A resistência cMLSB é facilmente detectada pelos testes de susceptibilidade utilizados na rotina laboratorial, mas a identificação da resistência iMLSB só é possível, mediante a utilização de um agente indutor. Dessa forma, a terapia com clindamicina nos casos de infecção por isolados com resistência iMLSB pode falhar. Assim, o presente estudo objetivou caracterizar o perfil fenotípico (cMLSB e iMLSB)e molecular (presença dos genes ermA, ermB, ermC e msrA) de 140 isolados clínicos de S. aureus sensíveis (MSSA) e resistentes à meticilina (MRSA), provenientes de pacientes admitidos no Hospital Universitário de Santa Maria (HUSM) no período compreendido entre abril a dezembro de 2011. Como critérios de inclusão foram considerados todos os isolados identificados como S. aureus através de metodologias fenotípicas manuais (coloração de Gram, prova da catalase, coagulase, etc.) e automatizada (MicroScan® - Siemens), sendo excluídas do estudo amostras repetidas do mesmo paciente. O fenótipo MLSB foi detectado através do teste de indução por disco aproximação (D-teste), sendo que das 140 cepas, 25 apresentaram fenótipo cMLSB e 11 o fenótipo iMLSB. Dos isolados constitutivos, 20 eram MRSA e 5 MSSA, enquanto nos isolados induzíveis, 8 (5,8%) eram MSSA e 3 (2,1%) MRSA. A partir desses resultados, foi realizada a caracterização molecular através da técnica de Reação em Cadeia da Polimerase (PCR) de todos os isolados que expressaram no teste fenotípico a resistência MLSB. Entre estes, os genes ermA e ermC foram os prevalentes, sendo identificados em 11 e 15 dos isolados respectivamente. O gene ermB amplificou em 4 das 36 cepas, enquanto o msrA em 13, sendo que destas, 8 apresentavam associação com os genes erm pesquisados. Apesar do fenótipo iMLSB ter sido menos frequente que o cMLSB, a realização do D-teste é importante para detecção destes fenótipos, pois pode orientar condutas terapêuticas, contribuindo para melhora na saúde do paciente, reduzindo o tempo de internação e consequentemente, o ônus para o sistema de saúde. Além disso, o teste é de baixo custo e fácil de ser executado, quando comparado a técnica da PCR, que apesar da alta sensibilidade e especificidade, não é empregada na rotina de todos os laboratórios clínicos, devido aos elevados custos.Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqporUniversidade Federal de Santa MariaCentro de Ciências da SaúdePrograma de Pós-Graduação em Ciências FarmacêuticasUFSMBrasilAnálises Clínicas e ToxicológicasAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessStaphylococcus aureus resistente à meticilina (MRSA)Staphylococcus aureus sensível à meticilina (MSSA)ClindamicinaResistência bacterianaClindamycinMethicillin-resistant Staphylococcus aureus (MRSA)Methicillin-sensitive Staphylococcus aureus (MSSA)Bacterial resistanceCNPQ::CIENCIAS DA SAUDE::FARMACIACaracterização fenotípica e molecular da resistência a clindamicina em isolados clínicos de Staphylococcus aureus de um hospital terciárioinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisHorner, Rosmarihttp://lattes.cnpq.br/5907084134183708Krause, Luciana Maria Fontanarihttp://lattes.cnpq.br/9844890896121847Pedro, Fabio Lopeshttp://lattes.cnpq.br/3151589084035930http://lattes.cnpq.br/5938466840557613Bottega, Angelita40030000000560071ad1279-3ed8-4b45-b319-57e5e774dd144b7d6f71-9a26-4d62-b5a1-bf6f1193adf6a310dec0-114a-4a8c-836d-d08756d18299d7b4058e-66a6-498a-a190-046ed2c26619reponame:Biblioteca Digital de Teses e Dissertações do UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMLICENSElicense.txtlicense.txttext/plain; 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dc.title.por.fl_str_mv Caracterização fenotípica e molecular da resistência a clindamicina em isolados clínicos de Staphylococcus aureus de um hospital terciário
title Caracterização fenotípica e molecular da resistência a clindamicina em isolados clínicos de Staphylococcus aureus de um hospital terciário
spellingShingle Caracterização fenotípica e molecular da resistência a clindamicina em isolados clínicos de Staphylococcus aureus de um hospital terciário
Bottega, Angelita
Staphylococcus aureus resistente à meticilina (MRSA)
Staphylococcus aureus sensível à meticilina (MSSA)
Clindamicina
Resistência bacteriana
Clindamycin
Methicillin-resistant Staphylococcus aureus (MRSA)
Methicillin-sensitive Staphylococcus aureus (MSSA)
Bacterial resistance
CNPQ::CIENCIAS DA SAUDE::FARMACIA
title_short Caracterização fenotípica e molecular da resistência a clindamicina em isolados clínicos de Staphylococcus aureus de um hospital terciário
title_full Caracterização fenotípica e molecular da resistência a clindamicina em isolados clínicos de Staphylococcus aureus de um hospital terciário
title_fullStr Caracterização fenotípica e molecular da resistência a clindamicina em isolados clínicos de Staphylococcus aureus de um hospital terciário
title_full_unstemmed Caracterização fenotípica e molecular da resistência a clindamicina em isolados clínicos de Staphylococcus aureus de um hospital terciário
title_sort Caracterização fenotípica e molecular da resistência a clindamicina em isolados clínicos de Staphylococcus aureus de um hospital terciário
author Bottega, Angelita
author_facet Bottega, Angelita
author_role author
dc.contributor.advisor1.fl_str_mv Horner, Rosmari
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/5907084134183708
dc.contributor.referee1.fl_str_mv Krause, Luciana Maria Fontanari
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/9844890896121847
dc.contributor.referee2.fl_str_mv Pedro, Fabio Lopes
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/3151589084035930
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/5938466840557613
dc.contributor.author.fl_str_mv Bottega, Angelita
contributor_str_mv Horner, Rosmari
Krause, Luciana Maria Fontanari
Pedro, Fabio Lopes
dc.subject.por.fl_str_mv Staphylococcus aureus resistente à meticilina (MRSA)
Staphylococcus aureus sensível à meticilina (MSSA)
Clindamicina
Resistência bacteriana
Clindamycin
topic Staphylococcus aureus resistente à meticilina (MRSA)
Staphylococcus aureus sensível à meticilina (MSSA)
Clindamicina
Resistência bacteriana
Clindamycin
Methicillin-resistant Staphylococcus aureus (MRSA)
Methicillin-sensitive Staphylococcus aureus (MSSA)
Bacterial resistance
CNPQ::CIENCIAS DA SAUDE::FARMACIA
dc.subject.eng.fl_str_mv Methicillin-resistant Staphylococcus aureus (MRSA)
Methicillin-sensitive Staphylococcus aureus (MSSA)
Bacterial resistance
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS DA SAUDE::FARMACIA
description Staphylococcus aureus demonstrated over the time, the ability to develop resistance to the most antimicrobials. The mechanism of resistance to macrolides in S. aureus also reaches lincosamides and streptogramins B, featuring the designate MLSB resistance, whose expression can be constitutive (cMLSB) or inducible (iMLSB) and it is encoded mainly by the presence of erm genes (ermA, ermB and ermC) and msrA. The cMLSB resistance is easily detected by susceptibility testing used in routine laboratory, but iMLSB resistance identification is only possible by using an inducing agent. Thus, clindamycin therapy in cases of infection by isolated with iMLSB resistance can fail. Thus, this study aimed to characterize the phenotypic profile (cMLSB e iMLSB) and molecular (presence of ermA, ermB, ermC and mrsA genes) of 140 clinical isolates of methicillin-sensitive S. aureus (MSSA) and methicillin-resistant (MRSA), from patients admitted at University Hospital of Santa Maria (HUSM) in the period from April to December 2011.The inclusion criteria were all isolates identified as S. aureus by manual phenotypic methods (Gram stain, catalase test, coagulase, etc.) and automated (MicroScan® - Siemens), and excluded of the study repeated samples from the same patient. The MLSB phenotype was detected by approaching disk induction test (D-test), and from 140 strains, 25 had cMLSB phenotype and 11 the iMLSB phenotype. Of constitutive isolates, 20 were MRSA and 5 MSSA, while the inducible isolates 8 (5.8%) were MSSA and 3 (2.1%) MRSA. From these results, we performed the molecular characterization by Reaction technique Polymerase Chain (PCR) of all isolates that expressed the MLSB resistance in phenotypic test. Among these, ermA and ermC genes were prevalent, identified at 11 and 15 of the isolates, respectively. The gene ermB amplified in 4of 36 strains, while msrA in 13, and of these, 8 had an association with the erm genes surveyed. Despite the iMLSB phenotype have been less frequent than cMLSB, the realization of the D-test is important to detect these phenotypes, it may guide therapeutic procedures, helping to improve in the health of the patient, reducing the length of hospitalization and thus the burden on the health system. Moreover, the test is inexpensive and easy to run compared to the PCR technique, which despite its high sensitivity and specificity is not used in routine of all clinical laboratories due to high costs.
publishDate 2016
dc.date.issued.fl_str_mv 2016-08-11
dc.date.accessioned.fl_str_mv 2019-09-06T21:34:10Z
dc.date.available.fl_str_mv 2019-09-06T21:34:10Z
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language por
dc.relation.cnpq.fl_str_mv 400300000005
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rights_invalid_str_mv Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Universidade Federal de Santa Maria
Centro de Ciências da Saúde
dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Ciências Farmacêuticas
dc.publisher.initials.fl_str_mv UFSM
dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Análises Clínicas e Toxicológicas
publisher.none.fl_str_mv Universidade Federal de Santa Maria
Centro de Ciências da Saúde
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