Cargando…
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...
Autores principales: | , , , , , , |
---|---|
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 |
_version_ | 1783235186706612224 |
---|---|
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. |
format | Online Article Text |
id | pubmed-5424758 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT mouramelosuely aquantitativepcrelectrochemicalgenosensortestforthescreeningofbiotechcrops AT mirandacastrorebeca aquantitativepcrelectrochemicalgenosensortestforthescreeningofbiotechcrops AT delossantosalvareznoemi aquantitativepcrelectrochemicalgenosensortestforthescreeningofbiotechcrops AT mirandaordieresarturoj aquantitativepcrelectrochemicalgenosensortestforthescreeningofbiotechcrops AT dossantosjuniorjoseribeiro aquantitativepcrelectrochemicalgenosensortestforthescreeningofbiotechcrops AT dasilvafonsecarosanaa aquantitativepcrelectrochemicalgenosensortestforthescreeningofbiotechcrops AT lobocastanonmariajesus aquantitativepcrelectrochemicalgenosensortestforthescreeningofbiotechcrops AT mouramelosuely quantitativepcrelectrochemicalgenosensortestforthescreeningofbiotechcrops AT mirandacastrorebeca quantitativepcrelectrochemicalgenosensortestforthescreeningofbiotechcrops AT delossantosalvareznoemi quantitativepcrelectrochemicalgenosensortestforthescreeningofbiotechcrops AT mirandaordieresarturoj quantitativepcrelectrochemicalgenosensortestforthescreeningofbiotechcrops AT dossantosjuniorjoseribeiro quantitativepcrelectrochemicalgenosensortestforthescreeningofbiotechcrops AT dasilvafonsecarosanaa quantitativepcrelectrochemicalgenosensortestforthescreeningofbiotechcrops AT lobocastanonmariajesus quantitativepcrelectrochemicalgenosensortestforthescreeningofbiotechcrops |