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Please use this identifier to cite or link to this item: http://hdl.handle.net/1843/41441
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dc.creatorCarolina Sheng Whei Miawpt_BR
dc.creatorEmanuel Novaes Vasconcelospt_BR
dc.creatorNilson César Castanheira Guimarãespt_BR
dc.creatorScheilla Vitorino Carvalho de Souzapt_BR
dc.date.accessioned2022-05-06T22:08:07Z-
dc.date.available2022-05-06T22:08:07Z-
dc.date.issued2017-
dc.citation.volume9pt_BR
dc.citation.issue4pt_BR
dc.citation.spage401pt_BR
dc.citation.epage412pt_BR
dc.identifier.doi10.3920/qas2016.0974pt_BR
dc.identifier.issn1757-8361pt_BR
dc.identifier.urihttp://hdl.handle.net/1843/41441-
dc.description.resumoThe 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.pt_BR
dc.description.sponsorshipCNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológicopt_BR
dc.format.mimetypepdfpt_BR
dc.languageengpt_BR
dc.publisherUniversidade Federal de Minas Geraispt_BR
dc.publisher.countryBrasilpt_BR
dc.publisher.departmentFAR - DEPARTAMENTO DE ALIMENTOSpt_BR
dc.publisher.initialsUFMGpt_BR
dc.relation.ispartofQuality Assurance and Safety of Crops & Foodspt_BR
dc.rightsAcesso Abertopt_BR
dc.subjectqualitative validationpt_BR
dc.subjectgenetically modified organismspt_BR
dc.subjectBt11 maizept_BR
dc.subjectreal time PCRpt_BR
dc.subjectDNA extractionpt_BR
dc.subjectDNA quantificationpt_BR
dc.subject.otherTecnologia de alimentospt_BR
dc.subject.otherMilhopt_BR
dc.subject.otherDNApt_BR
dc.subject.otherOrganismos geneticamente modificadospt_BR
dc.titleBt11 event detection by real-time PCR: single-laboratory validation, comparison of DNA extraction and quantification techniques and applicationpt_BR
dc.typeArtigo de Periódicopt_BR
dc.url.externahttps://qascf.com/index.php/qas/article/view/115pt_BR
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