Detection and quantification of Roundup Ready® soybean residues in sausage samples by conventional and real-time PCR

Detalhes bibliográficos
Autor(a) principal: De-Barros, Everaldo Gonçalves
Data de Publicação: 2008
Outros Autores: Marcelino, Francismar Corrêa, Guimarães, Marta Fonseca Martins
Tipo de documento: Artigo
Idioma: eng
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: http://dx.doi.org/10.1590/S0101-20612008000500007
http://locus.ufv.br//handle/123456789/25971
Resumo: The increasing presence of products derived from genetically modified (GM) plants in human and animal diets has led to the development of detection methods to distinguish biotechnology-derived foods from conventional ones. The conventional and real-time PCR have been used, respectively, to detect and quantify GM residues in highly processed foods. DNA extraction is a critical step during the analysis process. Some factors such as DNA degradation, matrix effects, and the presence of PCR inhibitors imply that a detection or quantification limit, established for a given method, is restricted to a matrix used during validation and cannot be projected to any other matrix outside the scope of the method. In Brazil, sausage samples were the main class of processed products in which Roundup Ready® (RR) soybean residues were detected. Thus, the validation of methodologies for the detection and quantification of those residues is absolutely necessary. Sausage samples were submitted to two different methods of DNA extraction: modified Wizard and the CTAB method. The yield and quality were compared for both methods. DNA samples were analyzed by conventional and real-time PCR for the detection and quantification of Roundup Ready® soybean in the samples. At least 200 ng of total sausage DNA was necessary for a reliable quantification. Reactions containing DNA amounts below this value led to large variations on the expected GM percentage value. In conventional PCR, the detection limit varied from 1.0 to 500 ng, depending on the GM soybean content in the sample. The precision, performance, and linearity were relatively high indicating that the method used for analysis was satisfactory.
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spelling Detection and quantification of Roundup Ready® soybean residues in sausage samples by conventional and real-time PCRPCR quantitativeGMOSausageTransgenic residuesPCR quantitativoOGMSalsichaResíduos transgênicosThe increasing presence of products derived from genetically modified (GM) plants in human and animal diets has led to the development of detection methods to distinguish biotechnology-derived foods from conventional ones. The conventional and real-time PCR have been used, respectively, to detect and quantify GM residues in highly processed foods. DNA extraction is a critical step during the analysis process. Some factors such as DNA degradation, matrix effects, and the presence of PCR inhibitors imply that a detection or quantification limit, established for a given method, is restricted to a matrix used during validation and cannot be projected to any other matrix outside the scope of the method. In Brazil, sausage samples were the main class of processed products in which Roundup Ready® (RR) soybean residues were detected. Thus, the validation of methodologies for the detection and quantification of those residues is absolutely necessary. Sausage samples were submitted to two different methods of DNA extraction: modified Wizard and the CTAB method. The yield and quality were compared for both methods. DNA samples were analyzed by conventional and real-time PCR for the detection and quantification of Roundup Ready® soybean in the samples. At least 200 ng of total sausage DNA was necessary for a reliable quantification. Reactions containing DNA amounts below this value led to large variations on the expected GM percentage value. In conventional PCR, the detection limit varied from 1.0 to 500 ng, depending on the GM soybean content in the sample. The precision, performance, and linearity were relatively high indicating that the method used for analysis was satisfactory.A presença crescente de produtos derivados de plantas geneticamente modificadas (GM) na dieta humana e de animais tem levado ao desenvolvimento de métodos de detecção capazes de distinguir entre alimentos derivados da biotecnologia e alimentos convencionais. As técnicas de PCR convencional e em tempo real têm sido as mais utilizadas para detectar e quantificar resíduos GM em alimentos altamente processados, respectivamente. A extração de DNA é um passo crítico durante o processo de análise. Alguns fatores, tais como, degradação de DNA, efeito de matriz e a presença de inibidores da PCR, implicam que um limite de detecção ou quantificação estabelecido para um determinado método é restrito à matriz usada durante validação e não pode ser estendido a qualquer outra matriz fora do escopo do método. No Brasil, amostras de salsicha constituíram a principal classe de produtos, nas quais resíduos de soja Roundup Ready® (RR) foram detectados. Então a validação de metodologias para detecção e quantificação destes resíduos é urgente. Amostras de salsicha foram submetidas a dois diferentes métodos de extração de DNA: Wizard modificado e CTAB. O rendimento e a qualidade foram comparados em ambos os métodos. As amostras foram analisadas por PCR convencional e em tempo real para detecção e quantificação de resíduos de soja RR. Pelo menos 200 ng de DNA total da amostra de salsicha foram necessários para garantir uma quantificação confiável. Reações contendo concentrações de DNA abaixo deste limite apresentaram grande variação nos valores de porcentagem de GM esperados. Na PCR convencional, o limite de detecção variou de 1,0 a 500 ng, dependendo do conteúdo de soja GM na amostra. A precisão, performance e linearidade foram relativamente altas, indicando que o método de análise foi satisfatório.Food Science and Technology2019-06-28T12:27:21Z2019-06-28T12:27:21Z2008-12info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlepdfapplication/pdf1678457Xhttp://dx.doi.org/10.1590/S0101-20612008000500007http://locus.ufv.br//handle/123456789/25971engv. 28, p. 38- 45, dez. 2008De-Barros, Everaldo GonçalvesMarcelino, Francismar CorrêaGuimarães, Marta Fonseca Martinsinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFV2024-07-12T08:07:36Zoai:locus.ufv.br:123456789/25971Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452024-07-12T08:07:36LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.none.fl_str_mv Detection and quantification of Roundup Ready® soybean residues in sausage samples by conventional and real-time PCR
title Detection and quantification of Roundup Ready® soybean residues in sausage samples by conventional and real-time PCR
spellingShingle Detection and quantification of Roundup Ready® soybean residues in sausage samples by conventional and real-time PCR
De-Barros, Everaldo Gonçalves
PCR quantitative
GMO
Sausage
Transgenic residues
PCR quantitativo
OGM
Salsicha
Resíduos transgênicos
title_short Detection and quantification of Roundup Ready® soybean residues in sausage samples by conventional and real-time PCR
title_full Detection and quantification of Roundup Ready® soybean residues in sausage samples by conventional and real-time PCR
title_fullStr Detection and quantification of Roundup Ready® soybean residues in sausage samples by conventional and real-time PCR
title_full_unstemmed Detection and quantification of Roundup Ready® soybean residues in sausage samples by conventional and real-time PCR
title_sort Detection and quantification of Roundup Ready® soybean residues in sausage samples by conventional and real-time PCR
author De-Barros, Everaldo Gonçalves
author_facet De-Barros, Everaldo Gonçalves
Marcelino, Francismar Corrêa
Guimarães, Marta Fonseca Martins
author_role author
author2 Marcelino, Francismar Corrêa
Guimarães, Marta Fonseca Martins
author2_role author
author
dc.contributor.author.fl_str_mv De-Barros, Everaldo Gonçalves
Marcelino, Francismar Corrêa
Guimarães, Marta Fonseca Martins
dc.subject.por.fl_str_mv PCR quantitative
GMO
Sausage
Transgenic residues
PCR quantitativo
OGM
Salsicha
Resíduos transgênicos
topic PCR quantitative
GMO
Sausage
Transgenic residues
PCR quantitativo
OGM
Salsicha
Resíduos transgênicos
description The increasing presence of products derived from genetically modified (GM) plants in human and animal diets has led to the development of detection methods to distinguish biotechnology-derived foods from conventional ones. The conventional and real-time PCR have been used, respectively, to detect and quantify GM residues in highly processed foods. DNA extraction is a critical step during the analysis process. Some factors such as DNA degradation, matrix effects, and the presence of PCR inhibitors imply that a detection or quantification limit, established for a given method, is restricted to a matrix used during validation and cannot be projected to any other matrix outside the scope of the method. In Brazil, sausage samples were the main class of processed products in which Roundup Ready® (RR) soybean residues were detected. Thus, the validation of methodologies for the detection and quantification of those residues is absolutely necessary. Sausage samples were submitted to two different methods of DNA extraction: modified Wizard and the CTAB method. The yield and quality were compared for both methods. DNA samples were analyzed by conventional and real-time PCR for the detection and quantification of Roundup Ready® soybean in the samples. At least 200 ng of total sausage DNA was necessary for a reliable quantification. Reactions containing DNA amounts below this value led to large variations on the expected GM percentage value. In conventional PCR, the detection limit varied from 1.0 to 500 ng, depending on the GM soybean content in the sample. The precision, performance, and linearity were relatively high indicating that the method used for analysis was satisfactory.
publishDate 2008
dc.date.none.fl_str_mv 2008-12
2019-06-28T12:27:21Z
2019-06-28T12:27:21Z
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 1678457X
http://dx.doi.org/10.1590/S0101-20612008000500007
http://locus.ufv.br//handle/123456789/25971
identifier_str_mv 1678457X
url http://dx.doi.org/10.1590/S0101-20612008000500007
http://locus.ufv.br//handle/123456789/25971
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv v. 28, p. 38- 45, dez. 2008
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv pdf
application/pdf
dc.publisher.none.fl_str_mv Food Science and Technology
publisher.none.fl_str_mv Food Science and Technology
dc.source.none.fl_str_mv reponame:LOCUS Repositório Institucional da UFV
instname:Universidade Federal de Viçosa (UFV)
instacron:UFV
instname_str Universidade Federal de Viçosa (UFV)
instacron_str UFV
institution UFV
reponame_str LOCUS Repositório Institucional da UFV
collection LOCUS Repositório Institucional da UFV
repository.name.fl_str_mv LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)
repository.mail.fl_str_mv fabiojreis@ufv.br
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