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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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Detalles Bibliográficos
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
Descripción
Sumario: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.