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A Quantitative PCR-Electrochemical Genosensor Test for the Screening of Biotech Crops

The design of screening methods for the detection of genetically modified organisms (GMOs) in food would improve the efficiency in their control. We report here a PCR amplification method combined with a sequence-specific electrochemical genosensor for the quantification of a DNA sequence characteri...

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Autores principales: Moura-Melo, Suely, Miranda-Castro, Rebeca, de-los-Santos-Álvarez, Noemí, Miranda-Ordieres, Arturo J., dos Santos Junior, José Ribeiro, da Silva Fonseca, Rosana A., Lobo-Castañón, María Jesús
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5424758/
https://www.ncbi.nlm.nih.gov/pubmed/28420193
http://dx.doi.org/10.3390/s17040881
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author Moura-Melo, Suely
Miranda-Castro, Rebeca
de-los-Santos-Álvarez, Noemí
Miranda-Ordieres, Arturo J.
dos Santos Junior, José Ribeiro
da Silva Fonseca, Rosana A.
Lobo-Castañón, María Jesús
author_facet Moura-Melo, Suely
Miranda-Castro, Rebeca
de-los-Santos-Álvarez, Noemí
Miranda-Ordieres, Arturo J.
dos Santos Junior, José Ribeiro
da Silva Fonseca, Rosana A.
Lobo-Castañón, María Jesús
author_sort Moura-Melo, Suely
collection PubMed
description The design of screening methods for the detection of genetically modified organisms (GMOs) in food would improve the efficiency in their control. We report here a PCR amplification method combined with a sequence-specific electrochemical genosensor for the quantification of a DNA sequence characteristic of the 35S promoter derived from the cauliflower mosaic virus (CaMV). Specifically, we employ a genosensor constructed by chemisorption of a thiolated capture probe and p-aminothiophenol gold surfaces to entrap on the sensing layer the unpurified PCR amplicons, together with a signaling probe labeled with fluorescein. The proposed test allows for the determination of a transgene copy number in both hemizygous (maize MON810 trait) and homozygous (soybean GTS40-3-2) transformed plants, and exhibits a limit of quantification of at least 0.25% for both kinds of GMO lines.
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spelling pubmed-54247582017-05-12 A Quantitative PCR-Electrochemical Genosensor Test for the Screening of Biotech Crops Moura-Melo, Suely Miranda-Castro, Rebeca de-los-Santos-Álvarez, Noemí Miranda-Ordieres, Arturo J. dos Santos Junior, José Ribeiro da Silva Fonseca, Rosana A. Lobo-Castañón, María Jesús Sensors (Basel) Article The design of screening methods for the detection of genetically modified organisms (GMOs) in food would improve the efficiency in their control. We report here a PCR amplification method combined with a sequence-specific electrochemical genosensor for the quantification of a DNA sequence characteristic of the 35S promoter derived from the cauliflower mosaic virus (CaMV). Specifically, we employ a genosensor constructed by chemisorption of a thiolated capture probe and p-aminothiophenol gold surfaces to entrap on the sensing layer the unpurified PCR amplicons, together with a signaling probe labeled with fluorescein. The proposed test allows for the determination of a transgene copy number in both hemizygous (maize MON810 trait) and homozygous (soybean GTS40-3-2) transformed plants, and exhibits a limit of quantification of at least 0.25% for both kinds of GMO lines. MDPI 2017-04-18 /pmc/articles/PMC5424758/ /pubmed/28420193 http://dx.doi.org/10.3390/s17040881 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Moura-Melo, Suely
Miranda-Castro, Rebeca
de-los-Santos-Álvarez, Noemí
Miranda-Ordieres, Arturo J.
dos Santos Junior, José Ribeiro
da Silva Fonseca, Rosana A.
Lobo-Castañón, María Jesús
A Quantitative PCR-Electrochemical Genosensor Test for the Screening of Biotech Crops
title A Quantitative PCR-Electrochemical Genosensor Test for the Screening of Biotech Crops
title_full A Quantitative PCR-Electrochemical Genosensor Test for the Screening of Biotech Crops
title_fullStr A Quantitative PCR-Electrochemical Genosensor Test for the Screening of Biotech Crops
title_full_unstemmed A Quantitative PCR-Electrochemical Genosensor Test for the Screening of Biotech Crops
title_short A Quantitative PCR-Electrochemical Genosensor Test for the Screening of Biotech Crops
title_sort quantitative pcr-electrochemical genosensor test for the screening of biotech crops
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5424758/
https://www.ncbi.nlm.nih.gov/pubmed/28420193
http://dx.doi.org/10.3390/s17040881
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