Bt11 event detection by real-time PCR: single-laboratory validation, comparison of DNA extraction and quantification techniques and application

Detalhes bibliográficos
Autor(a) principal: Carolina Sheng Whei Miaw
Data de Publicação: 2017
Outros Autores: Emanuel Novaes Vasconcelos, Nilson César Castanheira Guimarães, Scheilla Vitorino Carvalho de Souza
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UFMG
Texto Completo: http://hdl.handle.net/1843/41441
Resumo: CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico
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spelling 2022-05-06T22:08:07Z2022-05-06T22:08:07Z20179440141210.3920/qas2016.09741757-8361http://hdl.handle.net/1843/41441CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoThe detection of genetically modified organisms (GMO) by real-time polymerase chain reaction (PCR) is recommended due to its effectiveness in GMO analysis. A complete in-house validation method was applied to the detection of Bt11 events by real-time PCR. A full factorial design was used to compare DNA extraction (cetyltrimethyl ammonium bromide; CTAB, and NucleoSpin® Plant II Kit) and DNA quantification techniques (conventional GENESYS™ 10S UV-Vis spectrophotometer and confined drop-based NANOVUE™ Plus spectrophotometer). In the validation, various levels (0.0007 to 0.0315%) of Bt11 maize were formulated with blank maize and certified Bt11 reference material. A false-positive rate of 0% was obtained for blank samples, which corresponded to selectivity and reliability rates of 100%. The false-negative rate varied from 0 to 83.3%, consistent with sensitivity and reliability rates ranging from 16.7 to 100%. The Bt11 level that presented 100% positive results was 0.0315%, which indicated the sensitivity of the method. Non-linear models were used to estimate the region of unreliability and to calculate the detection limit of 0.014%. Accordance and concordance values of 1.0 were obtained for the 0.0315% level, which indicated method standardisation. Selectivity in the presence of interference was confirmed by the detection of Bt11 maize in the presence of other events. The method was considered robust for different DNA extraction and DNA quantification techniques. Higher DNA concentration values were obtained using CTAB. The absorbance ratio of A260/A230 was negatively influenced by quantification using a conventional spectrophotometer. Both DNA extraction techniques gave values of A260/A280 higher than 1.7, which indicated DNA of great purity. This validated method was applied to routine samples.engUniversidade Federal de Minas GeraisUFMGBrasilFAR - DEPARTAMENTO DE ALIMENTOSQuality Assurance and Safety of Crops & FoodsTecnologia de alimentosMilhoDNAOrganismos geneticamente modificadosqualitative validationgenetically modified organismsBt11 maizereal time PCRDNA extractionDNA quantificationBt11 event detection by real-time PCR: single-laboratory validation, comparison of DNA extraction and quantification techniques and applicationinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttps://qascf.com/index.php/qas/article/view/115Carolina Sheng Whei MiawEmanuel Novaes VasconcelosNilson César Castanheira GuimarãesScheilla Vitorino Carvalho de Souzaapplication/pdfinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGLICENSELicense.txtLicense.txttext/plain; charset=utf-82042https://repositorio.ufmg.br/bitstream/1843/41441/1/License.txtfa505098d172de0bc8864fc1287ffe22MD51ORIGINALBt11 event detection by real-time PCR single-laboratory validation, comparison of DNA extraction and quantification techniques and application.pdfBt11 event detection by real-time PCR single-laboratory validation, comparison of DNA extraction and quantification techniques and application.pdfapplication/pdf362147https://repositorio.ufmg.br/bitstream/1843/41441/2/Bt11%20event%20detection%20by%20real-time%20PCR%20single-laboratory%20validation%2c%20comparison%20of%20DNA%20extraction%20and%20quantification%20techniques%20and%20application.pdfebc9980cfdbb0016d374453bb515617fMD521843/414412022-05-06 19:08:07.543oai:repositorio.ufmg.br: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Repositório de PublicaçõesPUBhttps://repositorio.ufmg.br/oaiopendoar:2022-05-06T22:08:07Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
dc.title.pt_BR.fl_str_mv Bt11 event detection by real-time PCR: single-laboratory validation, comparison of DNA extraction and quantification techniques and application
title Bt11 event detection by real-time PCR: single-laboratory validation, comparison of DNA extraction and quantification techniques and application
spellingShingle Bt11 event detection by real-time PCR: single-laboratory validation, comparison of DNA extraction and quantification techniques and application
Carolina Sheng Whei Miaw
qualitative validation
genetically modified organisms
Bt11 maize
real time PCR
DNA extraction
DNA quantification
Tecnologia de alimentos
Milho
DNA
Organismos geneticamente modificados
title_short Bt11 event detection by real-time PCR: single-laboratory validation, comparison of DNA extraction and quantification techniques and application
title_full Bt11 event detection by real-time PCR: single-laboratory validation, comparison of DNA extraction and quantification techniques and application
title_fullStr Bt11 event detection by real-time PCR: single-laboratory validation, comparison of DNA extraction and quantification techniques and application
title_full_unstemmed Bt11 event detection by real-time PCR: single-laboratory validation, comparison of DNA extraction and quantification techniques and application
title_sort Bt11 event detection by real-time PCR: single-laboratory validation, comparison of DNA extraction and quantification techniques and application
author Carolina Sheng Whei Miaw
author_facet Carolina Sheng Whei Miaw
Emanuel Novaes Vasconcelos
Nilson César Castanheira Guimarães
Scheilla Vitorino Carvalho de Souza
author_role author
author2 Emanuel Novaes Vasconcelos
Nilson César Castanheira Guimarães
Scheilla Vitorino Carvalho de Souza
author2_role author
author
author
dc.contributor.author.fl_str_mv Carolina Sheng Whei Miaw
Emanuel Novaes Vasconcelos
Nilson César Castanheira Guimarães
Scheilla Vitorino Carvalho de Souza
dc.subject.por.fl_str_mv qualitative validation
genetically modified organisms
Bt11 maize
real time PCR
DNA extraction
DNA quantification
topic qualitative validation
genetically modified organisms
Bt11 maize
real time PCR
DNA extraction
DNA quantification
Tecnologia de alimentos
Milho
DNA
Organismos geneticamente modificados
dc.subject.other.pt_BR.fl_str_mv Tecnologia de alimentos
Milho
DNA
Organismos geneticamente modificados
description CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico
publishDate 2017
dc.date.issued.fl_str_mv 2017
dc.date.accessioned.fl_str_mv 2022-05-06T22:08:07Z
dc.date.available.fl_str_mv 2022-05-06T22:08:07Z
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://hdl.handle.net/1843/41441
dc.identifier.doi.pt_BR.fl_str_mv 10.3920/qas2016.0974
dc.identifier.issn.pt_BR.fl_str_mv 1757-8361
identifier_str_mv 10.3920/qas2016.0974
1757-8361
url http://hdl.handle.net/1843/41441
dc.language.iso.fl_str_mv eng
language eng
dc.relation.ispartof.pt_BR.fl_str_mv Quality Assurance and Safety of Crops & Foods
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Minas Gerais
dc.publisher.initials.fl_str_mv UFMG
dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv FAR - DEPARTAMENTO DE ALIMENTOS
publisher.none.fl_str_mv Universidade Federal de Minas Gerais
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFMG
instname:Universidade Federal de Minas Gerais (UFMG)
instacron:UFMG
instname_str Universidade Federal de Minas Gerais (UFMG)
instacron_str UFMG
institution UFMG
reponame_str Repositório Institucional da UFMG
collection Repositório Institucional da UFMG
bitstream.url.fl_str_mv https://repositorio.ufmg.br/bitstream/1843/41441/1/License.txt
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