COMPARISON OF DNA EXTRACTION METHODS IN POLYMERASE CHAIN REACTION FOR THE DETECTION OF Salmonella enterica SEROVAR ENTERITIDIS IN POULTRY PRODUCTS
Autor(a) principal: | |
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Data de Publicação: | 2011 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | por |
Título da fonte: | Ciência animal brasileira (Online) |
Texto Completo: | https://revistas.ufg.br/vet/article/view/3774 |
Resumo: | The identification of Salmonella spp. in food samples by microbiological diagnosis is time consuming, with approximately five different stages, requiring about 120 hours until the final result. The utilization of the polymerase chain reaction technique (PCR) can reduce this time, but substances present in samples may affect the reaction. The present work aimed to compare DNA extraction by thermic treatment and by the use of cetyltrimethil ammonium bromide (CTAB), in products originated from poultry houses corresponding to raw material (meat meal) and experimentally contaminated drag swabs. Materials obtained from the extractions were submitted to PCR, utilizing a pair of initiator oligonucleotides for amplification of Sdf 1 gene fragments. Comparing the methods of extraction, it was observed that when CTAB was employed, SE was detected in 70% of meat meal and in 80% of drag swabs, while the thermic treatment method yielded positive results in 20% of meat meal and in 40% of drag swabs. SE was detected under both methods utilized for DNA extraction, but the use of CTAB detected a greater number of positive samples, compared with thermal treatment.KEYWORDS: PCR, Salmonella Enteritidis, chicken, DNA extraction, CTAB. |
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COMPARISON OF DNA EXTRACTION METHODS IN POLYMERASE CHAIN REACTION FOR THE DETECTION OF Salmonella enterica SEROVAR ENTERITIDIS IN POULTRY PRODUCTSComparación de métodos para la extracción de ADN en la reacción en cadena de la polimerasa para detección de Salmonella enterica serotipo Enteritidis en productos avícolasCOMPARAÇÃO DE MÉTODOS PARA EXTRAÇÃO DE DNA NA REAÇÃO EM CADEIA DA POLIMERASE PARA DETECÇÃO DE Salmonella enterica SOROVAR Enteritidis EM PRODUTOS AVÍCOLASAnimal sanityPCRSalmonella Enteritidisaveextração de DNACTAB.The identification of Salmonella spp. in food samples by microbiological diagnosis is time consuming, with approximately five different stages, requiring about 120 hours until the final result. The utilization of the polymerase chain reaction technique (PCR) can reduce this time, but substances present in samples may affect the reaction. The present work aimed to compare DNA extraction by thermic treatment and by the use of cetyltrimethil ammonium bromide (CTAB), in products originated from poultry houses corresponding to raw material (meat meal) and experimentally contaminated drag swabs. Materials obtained from the extractions were submitted to PCR, utilizing a pair of initiator oligonucleotides for amplification of Sdf 1 gene fragments. Comparing the methods of extraction, it was observed that when CTAB was employed, SE was detected in 70% of meat meal and in 80% of drag swabs, while the thermic treatment method yielded positive results in 20% of meat meal and in 40% of drag swabs. SE was detected under both methods utilized for DNA extraction, but the use of CTAB detected a greater number of positive samples, compared with thermal treatment.KEYWORDS: PCR, Salmonella Enteritidis, chicken, DNA extraction, CTAB.La identificación de Salmonella spp. por diagnóstico microbiológico en muestras de alimentos es tardada, y consta de aproximadamente cinco etapas diferentes, lo que lleva cerca de 120 horas hasta el resultado final. La utilización de la técnica de reacción en cadena de la polimerasa (PCR) puede disminuir dicho periodo. No obstante, sustancias presentes en las muestras pueden afectar el resultado de la prueba. El objetivo de este trabajo fue comparar la extracción de ADN por tratamiento térmico y por el uso de bromuro de cetiltrimetil amonio (CTAB) en PCR para la detección de Salmonellla enterica serotipo Enteritidis (SE) en productos utilizados como materia prima provenientes de granjas avícolas (harina de carne) y frotes de arrastre contaminados experimentalmente. Los materiales obtenidos con las extracciones fueron sometidos a PCR, utilizando un par de oligonucleótidos iniciadores para la amplificación del ADN de lo gene Sdf 1. Basados en la comparación de los métodos de extracción, se observó que usando el CTBA se detectó SE en 70% de la materia prima y 80% en frotes de arrastre, mientras que con el método de tratamiento térmico hubo una positividad de la bacteria de 20% en materia prima y 40% en frotes de arrastre. Hubo detección de SE en los dos métodos para extracción de ADN, sin embargo, en comparación con el tratamiento térmico, el uso de CTBA detectó como positivas una mayor cantidad de muestras.A identificação de Salmonella spp. em amostras de alimentos por diagnóstico microbiológico é demorada, com aproximadamente cinco diferentes etapas, levando cerca de 120 horas até o resultado final. A utilização da técnica da reação em cadeia da polimerase (PCR) pode diminuir esse período; entretanto, substâncias presentes nas amostras podem afetar a reação. O objetivo deste trabalho foi comparar a extração de DNA de Salmonella enterica sorovar Enteritidis (SE), por tratamento térmico e pelo uso do cetyltrimethil ammonium bromide (CTAB), em produtos provenientes de granjas avícolas, correspondentes a matéria prima (farinha de carne) e a suabes de arrasto experimentalmente contaminados. As amostras de DNA obtidas com as extrações foram submetidas a PCR, utilizando-se um par de oligonucleotídeos iniciadores para amplificação de DNA do gene específico de SE – Sdf 1. Comparando-se os métodos de extração, observou-se que quando do uso do CTAB detectou-se SE em 70% de matéria prima e em 80% de suabes de arrasto, enquanto com o método por tratamento térmico houve positividade em 20% de matéria prima e em 40% de suabes de arrasto. Houve detecção de SE em ambos os métodos utilizados para extração de DNA, porém, o uso de CTAB detectou maior número de amostras positivas, quando comparado com o tratamento térmico.PALAVRAS-CHAVES: PCR, Salmonella Enteritidis, ave, extração de DNA, CTAB.Universidade Federal de Goiás2011-03-31info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionResearchAvaliado por paresPesquisa Científicaapplication/pdftext/htmlhttps://revistas.ufg.br/vet/article/view/377410.5216/cab.v12i1.3774Brazilian Animal Science/ Ciência Animal Brasileira; Vol. 12 No. 1 (2011); 115 - 119Ciência Animal Brasileira / Brazilian Animal Science; v. 12 n. 1 (2011); 115 - 1191809-68911518-2797reponame:Ciência animal brasileira (Online)instname:Universidade Federal de Goiás (UFG)instacron:UFGporhttps://revistas.ufg.br/vet/article/view/3774/9651https://revistas.ufg.br/vet/article/view/3774/8905Andreatti Filho, Raphael LucioMarietto Gonçalves, Guilherme AugustoOkamoto, Adriano SakaiLima, Edna Teresa deinfo:eu-repo/semantics/openAccess2011-04-22T14:34:11Zoai:ojs.revistas.ufg.br:article/3774Revistahttps://revistas.ufg.br/vetPUBhttps://revistas.ufg.br/vet/oai||revistacab@gmail.com1809-68911518-2797opendoar:2024-05-21T19:55:10.251111Ciência animal brasileira (Online) - Universidade Federal de Goiás (UFG)true |
dc.title.none.fl_str_mv |
COMPARISON OF DNA EXTRACTION METHODS IN POLYMERASE CHAIN REACTION FOR THE DETECTION OF Salmonella enterica SEROVAR ENTERITIDIS IN POULTRY PRODUCTS Comparación de métodos para la extracción de ADN en la reacción en cadena de la polimerasa para detección de Salmonella enterica serotipo Enteritidis en productos avícolas COMPARAÇÃO DE MÉTODOS PARA EXTRAÇÃO DE DNA NA REAÇÃO EM CADEIA DA POLIMERASE PARA DETECÇÃO DE Salmonella enterica SOROVAR Enteritidis EM PRODUTOS AVÍCOLAS |
title |
COMPARISON OF DNA EXTRACTION METHODS IN POLYMERASE CHAIN REACTION FOR THE DETECTION OF Salmonella enterica SEROVAR ENTERITIDIS IN POULTRY PRODUCTS |
spellingShingle |
COMPARISON OF DNA EXTRACTION METHODS IN POLYMERASE CHAIN REACTION FOR THE DETECTION OF Salmonella enterica SEROVAR ENTERITIDIS IN POULTRY PRODUCTS Andreatti Filho, Raphael Lucio Animal sanity PCR Salmonella Enteritidis ave extração de DNA CTAB. |
title_short |
COMPARISON OF DNA EXTRACTION METHODS IN POLYMERASE CHAIN REACTION FOR THE DETECTION OF Salmonella enterica SEROVAR ENTERITIDIS IN POULTRY PRODUCTS |
title_full |
COMPARISON OF DNA EXTRACTION METHODS IN POLYMERASE CHAIN REACTION FOR THE DETECTION OF Salmonella enterica SEROVAR ENTERITIDIS IN POULTRY PRODUCTS |
title_fullStr |
COMPARISON OF DNA EXTRACTION METHODS IN POLYMERASE CHAIN REACTION FOR THE DETECTION OF Salmonella enterica SEROVAR ENTERITIDIS IN POULTRY PRODUCTS |
title_full_unstemmed |
COMPARISON OF DNA EXTRACTION METHODS IN POLYMERASE CHAIN REACTION FOR THE DETECTION OF Salmonella enterica SEROVAR ENTERITIDIS IN POULTRY PRODUCTS |
title_sort |
COMPARISON OF DNA EXTRACTION METHODS IN POLYMERASE CHAIN REACTION FOR THE DETECTION OF Salmonella enterica SEROVAR ENTERITIDIS IN POULTRY PRODUCTS |
author |
Andreatti Filho, Raphael Lucio |
author_facet |
Andreatti Filho, Raphael Lucio Marietto Gonçalves, Guilherme Augusto Okamoto, Adriano Sakai Lima, Edna Teresa de |
author_role |
author |
author2 |
Marietto Gonçalves, Guilherme Augusto Okamoto, Adriano Sakai Lima, Edna Teresa de |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Andreatti Filho, Raphael Lucio Marietto Gonçalves, Guilherme Augusto Okamoto, Adriano Sakai Lima, Edna Teresa de |
dc.subject.por.fl_str_mv |
Animal sanity PCR Salmonella Enteritidis ave extração de DNA CTAB. |
topic |
Animal sanity PCR Salmonella Enteritidis ave extração de DNA CTAB. |
description |
The identification of Salmonella spp. in food samples by microbiological diagnosis is time consuming, with approximately five different stages, requiring about 120 hours until the final result. The utilization of the polymerase chain reaction technique (PCR) can reduce this time, but substances present in samples may affect the reaction. The present work aimed to compare DNA extraction by thermic treatment and by the use of cetyltrimethil ammonium bromide (CTAB), in products originated from poultry houses corresponding to raw material (meat meal) and experimentally contaminated drag swabs. Materials obtained from the extractions were submitted to PCR, utilizing a pair of initiator oligonucleotides for amplification of Sdf 1 gene fragments. Comparing the methods of extraction, it was observed that when CTAB was employed, SE was detected in 70% of meat meal and in 80% of drag swabs, while the thermic treatment method yielded positive results in 20% of meat meal and in 40% of drag swabs. SE was detected under both methods utilized for DNA extraction, but the use of CTAB detected a greater number of positive samples, compared with thermal treatment.KEYWORDS: PCR, Salmonella Enteritidis, chicken, DNA extraction, CTAB. |
publishDate |
2011 |
dc.date.none.fl_str_mv |
2011-03-31 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Research Avaliado por pares Pesquisa Científica |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://revistas.ufg.br/vet/article/view/3774 10.5216/cab.v12i1.3774 |
url |
https://revistas.ufg.br/vet/article/view/3774 |
identifier_str_mv |
10.5216/cab.v12i1.3774 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.none.fl_str_mv |
https://revistas.ufg.br/vet/article/view/3774/9651 https://revistas.ufg.br/vet/article/view/3774/8905 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf text/html |
dc.publisher.none.fl_str_mv |
Universidade Federal de Goiás |
publisher.none.fl_str_mv |
Universidade Federal de Goiás |
dc.source.none.fl_str_mv |
Brazilian Animal Science/ Ciência Animal Brasileira; Vol. 12 No. 1 (2011); 115 - 119 Ciência Animal Brasileira / Brazilian Animal Science; v. 12 n. 1 (2011); 115 - 119 1809-6891 1518-2797 reponame:Ciência animal brasileira (Online) instname:Universidade Federal de Goiás (UFG) instacron:UFG |
instname_str |
Universidade Federal de Goiás (UFG) |
instacron_str |
UFG |
institution |
UFG |
reponame_str |
Ciência animal brasileira (Online) |
collection |
Ciência animal brasileira (Online) |
repository.name.fl_str_mv |
Ciência animal brasileira (Online) - Universidade Federal de Goiás (UFG) |
repository.mail.fl_str_mv |
||revistacab@gmail.com |
_version_ |
1799874784526336000 |