COMPARISON OF DNA EXTRACTION METHODS IN POLYMERASE CHAIN REACTION FOR THE DETECTION OF Salmonella enterica SEROVAR ENTERITIDIS IN POULTRY PRODUCTS

Detalhes bibliográficos
Autor(a) principal: Andreatti Filho, Raphael Lucio
Data de Publicação: 2011
Outros Autores: Marietto Gonçalves, Guilherme Augusto, Okamoto, Adriano Sakai, Lima, Edna Teresa de
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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spelling 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
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