Multiplex PCR use for Staphylococcus aureus identification and oxacillin and mupirocin resistance evaluation

Detalhes bibliográficos
Autor(a) principal: Braoios, Alexandre
Data de Publicação: 2009
Outros Autores: Fluminhan, A., Pizzolitto, Antonio Carlos [UNESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://serv-bib.fcfar.unesp.br/seer/index.php/Cien_Farm/article/view/619
http://hdl.handle.net/11449/71141
Resumo: Oxacillin-resistant Staphylococcus aureus represents a serious problem in hospitals worldwide, increasing infected patients' mortality and morbidity and raising treatment costs and internment time. In this study, the results of using the Multiplex PCR technique to amplify fragments of the genes femA (specific-species), mecA (oxacillin resistance) and ileS-2 (mupirocin resistance) were compared with those of tests conventionally used to identify S. aureus isolates and ascertain their resistance to drugs. Fifty S. aureus strains were isolated from patients receiving treatment at UNOESTE University Hospital in Presidente Prudente, SP, Brazil. The 686 bp fragment corresponding to the gene femA was amplified and detected in all the isolates. On the other hand, the 310 bp fragment corresponding to the mecA gene was amplified in 29 (58%) of the isolates. All of the isolates showed sensitivity to mupirocin in the agar diffusion test, which was corroborated by the lack of any amplicon of the 456 bp fragment corresponding to the ileS-2 gene, in the PCR bands. The conventional tests to identify S. aureus and detect resistance to oxacillin and mupirocin showed 100% agreement with the PCR Multiplex results. The use of techniques for rapid and accurate identification of bacteria and assessment of their resistance may be valuable in the control of infection by resistant strains, allowing the rapid isolation and treatment of an infected patient. However, the results demonstrate that traditional phenotypic tests are also reliable, though they take more time.
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spelling Multiplex PCR use for Staphylococcus aureus identification and oxacillin and mupirocin resistance evaluationMupirocinOxacillin-resistantPCR multiplexStaphylococcus aureusbacterial proteinfemA proteinileS 2 proteinoxacillinpenicillin binding protein 2apseudomonic acidunclassified drugaccuracyagar diffusionampliconantibiotic resistancebacterial genebacterial strainbacterium identificationbacterium isolatebacterium isolationBrazilcontrolled studygene amplificationmultiplex polymerase chain reactionnonhumanreliabilityOxacillin-resistant Staphylococcus aureus represents a serious problem in hospitals worldwide, increasing infected patients' mortality and morbidity and raising treatment costs and internment time. In this study, the results of using the Multiplex PCR technique to amplify fragments of the genes femA (specific-species), mecA (oxacillin resistance) and ileS-2 (mupirocin resistance) were compared with those of tests conventionally used to identify S. aureus isolates and ascertain their resistance to drugs. Fifty S. aureus strains were isolated from patients receiving treatment at UNOESTE University Hospital in Presidente Prudente, SP, Brazil. The 686 bp fragment corresponding to the gene femA was amplified and detected in all the isolates. On the other hand, the 310 bp fragment corresponding to the mecA gene was amplified in 29 (58%) of the isolates. All of the isolates showed sensitivity to mupirocin in the agar diffusion test, which was corroborated by the lack of any amplicon of the 456 bp fragment corresponding to the ileS-2 gene, in the PCR bands. The conventional tests to identify S. aureus and detect resistance to oxacillin and mupirocin showed 100% agreement with the PCR Multiplex results. The use of techniques for rapid and accurate identification of bacteria and assessment of their resistance may be valuable in the control of infection by resistant strains, allowing the rapid isolation and treatment of an infected patient. However, the results demonstrate that traditional phenotypic tests are also reliable, though they take more time.Utilização de PCR-multiplex para identificação de Staphylococcus aureus e avaliação da resistência à oxacilina e mupirocina Staphylococcus aureus resistente à oxacilina representa um problema grave em hospitais de todo o mundo, aumentando a morbidade e mortalidade de pacientes infectados e, elevando os custos do tratamento e tempo de internação. Neste trabalho, foram comparados os resultados da técnica de PCR Multiplex para amplificação dos fragmentos dos genes femA (espécie-específico), mecA (resistência à oxacilina) e ileS-2 (resistência à mupirocina) com os resultados da identificação e testes convencionais para avaliação da resistência. Cinqüenta S. aureus foram isolados de pacientes atendidos no Hospital Universitário Dr. Domingos Leonardo Cerávolo da Unoeste, em Presidente Prudente, SP, Brasil. Houve amplificação do fragmento 686 pb, correspondente ao gene femA para todos os isolados. Por outro lado, houve amplificação do fragmento 310 pb correspondente ao gene mecA em 29 (58%) dos isolados. Todos os isolados mostraram sensibilidade à mupirocina observados no teste da difusão em ágar, e também pela ausência de amplificação do fragmento 456 pb, correspondente ao gene ileS-2. Os resultados dos testes convencionais para identificação de S. aureus e avaliação de resistência à oxacilina e mupirocina mostrou 100% de concordância com os resultados da PCR Multiplex. A utilização de técnicas mais rápidas e precisas para identificação e avaliação de resistência pode ser valiosa para o controle de infecção por cepas resistentes, permitindo rápido isolamento e tratamento do paciente. Entretanto os resultados demonstram que testes fenotípicos tradicionais também são confiáveis, apesar do maior tempo de execução. Palavras-chave: PCR Multiplex. Resistência à oxacilina. Staphylococcus aureus. Mupirocina.Departamento de Biomedicina Universidade Federal de Goiás - UGF, Jataí - GODepartamento de Genética Molecular Universidade do Oeste Paulista - UNOESTE, Presidente Prudente, SPDepartamento de Análises Clínicas Faculdade de Ciências Farmacêuticas UNESP, Araraquara, SPDepartamento de Análises Clínicas Faculdade de Ciências Farmacêuticas UNESP, Araraquara, SPUniversidade Federal de Goiás (UFG)Universidade do Oeste Paulista (UNOESTE)Universidade Estadual Paulista (Unesp)Braoios, AlexandreFluminhan, A.Pizzolitto, Antonio Carlos [UNESP]2014-05-27T11:23:58Z2014-05-27T11:23:58Z2009-09-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article303-307application/pdfhttp://serv-bib.fcfar.unesp.br/seer/index.php/Cien_Farm/article/view/619Revista de Ciencias Farmaceuticas Basica e Aplicada, v. 30, n. 3, p. 303-307, 2009.1808-4532http://hdl.handle.net/11449/711412-s2.0-786498499402-s2.0-78649849940.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengRevista de Ciências Farmacêuticas Básica e Aplicada0,131info:eu-repo/semantics/openAccess2024-06-21T15:18:46Zoai:repositorio.unesp.br:11449/71141Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T17:44:34.104682Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Multiplex PCR use for Staphylococcus aureus identification and oxacillin and mupirocin resistance evaluation
title Multiplex PCR use for Staphylococcus aureus identification and oxacillin and mupirocin resistance evaluation
spellingShingle Multiplex PCR use for Staphylococcus aureus identification and oxacillin and mupirocin resistance evaluation
Braoios, Alexandre
Mupirocin
Oxacillin-resistant
PCR multiplex
Staphylococcus aureus
bacterial protein
femA protein
ileS 2 protein
oxacillin
penicillin binding protein 2a
pseudomonic acid
unclassified drug
accuracy
agar diffusion
amplicon
antibiotic resistance
bacterial gene
bacterial strain
bacterium identification
bacterium isolate
bacterium isolation
Brazil
controlled study
gene amplification
multiplex polymerase chain reaction
nonhuman
reliability
title_short Multiplex PCR use for Staphylococcus aureus identification and oxacillin and mupirocin resistance evaluation
title_full Multiplex PCR use for Staphylococcus aureus identification and oxacillin and mupirocin resistance evaluation
title_fullStr Multiplex PCR use for Staphylococcus aureus identification and oxacillin and mupirocin resistance evaluation
title_full_unstemmed Multiplex PCR use for Staphylococcus aureus identification and oxacillin and mupirocin resistance evaluation
title_sort Multiplex PCR use for Staphylococcus aureus identification and oxacillin and mupirocin resistance evaluation
author Braoios, Alexandre
author_facet Braoios, Alexandre
Fluminhan, A.
Pizzolitto, Antonio Carlos [UNESP]
author_role author
author2 Fluminhan, A.
Pizzolitto, Antonio Carlos [UNESP]
author2_role author
author
dc.contributor.none.fl_str_mv Universidade Federal de Goiás (UFG)
Universidade do Oeste Paulista (UNOESTE)
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Braoios, Alexandre
Fluminhan, A.
Pizzolitto, Antonio Carlos [UNESP]
dc.subject.por.fl_str_mv Mupirocin
Oxacillin-resistant
PCR multiplex
Staphylococcus aureus
bacterial protein
femA protein
ileS 2 protein
oxacillin
penicillin binding protein 2a
pseudomonic acid
unclassified drug
accuracy
agar diffusion
amplicon
antibiotic resistance
bacterial gene
bacterial strain
bacterium identification
bacterium isolate
bacterium isolation
Brazil
controlled study
gene amplification
multiplex polymerase chain reaction
nonhuman
reliability
topic Mupirocin
Oxacillin-resistant
PCR multiplex
Staphylococcus aureus
bacterial protein
femA protein
ileS 2 protein
oxacillin
penicillin binding protein 2a
pseudomonic acid
unclassified drug
accuracy
agar diffusion
amplicon
antibiotic resistance
bacterial gene
bacterial strain
bacterium identification
bacterium isolate
bacterium isolation
Brazil
controlled study
gene amplification
multiplex polymerase chain reaction
nonhuman
reliability
description Oxacillin-resistant Staphylococcus aureus represents a serious problem in hospitals worldwide, increasing infected patients' mortality and morbidity and raising treatment costs and internment time. In this study, the results of using the Multiplex PCR technique to amplify fragments of the genes femA (specific-species), mecA (oxacillin resistance) and ileS-2 (mupirocin resistance) were compared with those of tests conventionally used to identify S. aureus isolates and ascertain their resistance to drugs. Fifty S. aureus strains were isolated from patients receiving treatment at UNOESTE University Hospital in Presidente Prudente, SP, Brazil. The 686 bp fragment corresponding to the gene femA was amplified and detected in all the isolates. On the other hand, the 310 bp fragment corresponding to the mecA gene was amplified in 29 (58%) of the isolates. All of the isolates showed sensitivity to mupirocin in the agar diffusion test, which was corroborated by the lack of any amplicon of the 456 bp fragment corresponding to the ileS-2 gene, in the PCR bands. The conventional tests to identify S. aureus and detect resistance to oxacillin and mupirocin showed 100% agreement with the PCR Multiplex results. The use of techniques for rapid and accurate identification of bacteria and assessment of their resistance may be valuable in the control of infection by resistant strains, allowing the rapid isolation and treatment of an infected patient. However, the results demonstrate that traditional phenotypic tests are also reliable, though they take more time.
publishDate 2009
dc.date.none.fl_str_mv 2009-09-01
2014-05-27T11:23:58Z
2014-05-27T11:23:58Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://serv-bib.fcfar.unesp.br/seer/index.php/Cien_Farm/article/view/619
Revista de Ciencias Farmaceuticas Basica e Aplicada, v. 30, n. 3, p. 303-307, 2009.
1808-4532
http://hdl.handle.net/11449/71141
2-s2.0-78649849940
2-s2.0-78649849940.pdf
url http://serv-bib.fcfar.unesp.br/seer/index.php/Cien_Farm/article/view/619
http://hdl.handle.net/11449/71141
identifier_str_mv Revista de Ciencias Farmaceuticas Basica e Aplicada, v. 30, n. 3, p. 303-307, 2009.
1808-4532
2-s2.0-78649849940
2-s2.0-78649849940.pdf
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Revista de Ciências Farmacêuticas Básica e Aplicada
0,131
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 303-307
application/pdf
dc.source.none.fl_str_mv Scopus
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
_version_ 1808128852839366656