New method for early detection of two random amplified polymorphic DNA (RAPD) groups of Staphylococcus aureus causing bovine mastitis infection in Paraná State, Brazil
The aim of this work was to develop a fast and accurate molecular approach to allow early detection of two RAPD groups of S. aureus causing bovine mastitis. Seventy five S. aureus isolates from infected animals were characterized by RAPD. Genomic fragments isolated from the unique bands present in e...
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Instituto de Tecnologia do Paraná (Tecpar)
2010-04-01
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doaj-835ac8ec1eb5462197e8fff973073a1b2020-11-24T23:23:49ZengInstituto de Tecnologia do Paraná (Tecpar)Brazilian Archives of Biology and Technology1516-89131678-43242010-04-0153235336010.1590/S1516-89132010000200014New method for early detection of two random amplified polymorphic DNA (RAPD) groups of Staphylococcus aureus causing bovine mastitis infection in Paraná State, BrazilDicezar GonçalvesJane Eire GabrielHumberto Maciel França MadeiraGuilherme Schnell e SchühliVânia Aparecida VicenteThe aim of this work was to develop a fast and accurate molecular approach to allow early detection of two RAPD groups of S. aureus causing bovine mastitis. Seventy five S. aureus isolates from infected animals were characterized by RAPD. Genomic fragments isolated from the unique bands present in either group were cloned and sequenced. Based on the DNA sequences, specific primers were designed to allow for the simultaneous detection of either group by multiplex PCR of S. aureus DNA isolated from clinical and subclinical bovine mastitis. Results showed that these proposed primers set could be used to detect various clinical and subclinical S. aureus isolates as well as the detection of the microorganism in bulk milk. Their use as a specific method for effective and early diagnostic tool for S. aureus infection in dairy herds is suggested.<br>Esta pesquisa objetivou o desenvolvimento de técnica rápida e eficiente para diagnosticar precocemente diferentes linhagens de S. aureus causadoras de mastite bovina. Como resultados da metodologia empregada, foram isoladas duas linhagens destas bactérias que causam diferentes tipos de mastite bovina. Os fragmentos de DNA genômico caracterizando ambas as linhagens, por meio de RAPD foram inseridos em vetor plasmidial pGEM e clonados por meio de clones T10 F1 de Escherichia coli. As seqüências obtidas permitiram desenhar iniciadores específicos para o reconhecimento de ambas as linhagens, os quais foram testados com amostras de S. aureus e com outras linhagens próximas. O diagnóstico por meios moleculares, pode ser realizado diretamente de amostras coletadas de rebanhos leiteiros assim como dos equipamentos de ordenha. A significância deste estudo consiste em um rápido e acurado método para localizar animais infectados, representando importante ferramenta no manejo do rebanho, na redução de custos com tratamentos e, rápida recuperação de rebanhos infectados.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132010000200014Staphylococcus aureusmolecular markerssubclinical mastitisclinical mastitismastitis detection |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Dicezar Gonçalves Jane Eire Gabriel Humberto Maciel França Madeira Guilherme Schnell e Schühli Vânia Aparecida Vicente |
spellingShingle |
Dicezar Gonçalves Jane Eire Gabriel Humberto Maciel França Madeira Guilherme Schnell e Schühli Vânia Aparecida Vicente New method for early detection of two random amplified polymorphic DNA (RAPD) groups of Staphylococcus aureus causing bovine mastitis infection in Paraná State, Brazil Brazilian Archives of Biology and Technology Staphylococcus aureus molecular markers subclinical mastitis clinical mastitis mastitis detection |
author_facet |
Dicezar Gonçalves Jane Eire Gabriel Humberto Maciel França Madeira Guilherme Schnell e Schühli Vânia Aparecida Vicente |
author_sort |
Dicezar Gonçalves |
title |
New method for early detection of two random amplified polymorphic DNA (RAPD) groups of Staphylococcus aureus causing bovine mastitis infection in Paraná State, Brazil |
title_short |
New method for early detection of two random amplified polymorphic DNA (RAPD) groups of Staphylococcus aureus causing bovine mastitis infection in Paraná State, Brazil |
title_full |
New method for early detection of two random amplified polymorphic DNA (RAPD) groups of Staphylococcus aureus causing bovine mastitis infection in Paraná State, Brazil |
title_fullStr |
New method for early detection of two random amplified polymorphic DNA (RAPD) groups of Staphylococcus aureus causing bovine mastitis infection in Paraná State, Brazil |
title_full_unstemmed |
New method for early detection of two random amplified polymorphic DNA (RAPD) groups of Staphylococcus aureus causing bovine mastitis infection in Paraná State, Brazil |
title_sort |
new method for early detection of two random amplified polymorphic dna (rapd) groups of staphylococcus aureus causing bovine mastitis infection in paraná state, brazil |
publisher |
Instituto de Tecnologia do Paraná (Tecpar) |
series |
Brazilian Archives of Biology and Technology |
issn |
1516-8913 1678-4324 |
publishDate |
2010-04-01 |
description |
The aim of this work was to develop a fast and accurate molecular approach to allow early detection of two RAPD groups of S. aureus causing bovine mastitis. Seventy five S. aureus isolates from infected animals were characterized by RAPD. Genomic fragments isolated from the unique bands present in either group were cloned and sequenced. Based on the DNA sequences, specific primers were designed to allow for the simultaneous detection of either group by multiplex PCR of S. aureus DNA isolated from clinical and subclinical bovine mastitis. Results showed that these proposed primers set could be used to detect various clinical and subclinical S. aureus isolates as well as the detection of the microorganism in bulk milk. Their use as a specific method for effective and early diagnostic tool for S. aureus infection in dairy herds is suggested.<br>Esta pesquisa objetivou o desenvolvimento de técnica rápida e eficiente para diagnosticar precocemente diferentes linhagens de S. aureus causadoras de mastite bovina. Como resultados da metodologia empregada, foram isoladas duas linhagens destas bactérias que causam diferentes tipos de mastite bovina. Os fragmentos de DNA genômico caracterizando ambas as linhagens, por meio de RAPD foram inseridos em vetor plasmidial pGEM e clonados por meio de clones T10 F1 de Escherichia coli. As seqüências obtidas permitiram desenhar iniciadores específicos para o reconhecimento de ambas as linhagens, os quais foram testados com amostras de S. aureus e com outras linhagens próximas. O diagnóstico por meios moleculares, pode ser realizado diretamente de amostras coletadas de rebanhos leiteiros assim como dos equipamentos de ordenha. A significância deste estudo consiste em um rápido e acurado método para localizar animais infectados, representando importante ferramenta no manejo do rebanho, na redução de custos com tratamentos e, rápida recuperação de rebanhos infectados. |
topic |
Staphylococcus aureus molecular markers subclinical mastitis clinical mastitis mastitis detection |
url |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132010000200014 |
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