Development of a Real-Time PCR Method for the Differential Detection and Quantification of Four Solanaceae in GMO Analysis: Potato (Solanum Tuberosum), Tomato (Solanum Lycopersicum), Eggplant (Solanum Melongena), and Pepper (Capsicum Annuum)
The labeling of products containing genetically modified organisms (GMO) is linked to their quantification since a threshold for the presence of fortuitous GMOs in food has been established. This threshold is calculated from a combination of two absolute quantification values: one for the specific G...
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Published in | Journal of agricultural and food chemistry Vol. 56; no. 6; pp. 1818 - 1828 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
Washington, DC
American Chemical Society
26.03.2008
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Subjects | |
Online Access | Get full text |
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Summary: | The labeling of products containing genetically modified organisms (GMO) is linked to their quantification since a threshold for the presence of fortuitous GMOs in food has been established. This threshold is calculated from a combination of two absolute quantification values: one for the specific GMO target and the second for an endogenous reference gene specific to the taxon. Thus, the development of reliable methods to quantify GMOs using endogenous reference genes in complex matrixes such as food and feed is needed. Plant identification can be difficult in the case of closely related taxa, which moreover are subject to introgression events. Based on the homology of β-fructosidase sequences obtained from public databases, two couples of consensus primers were designed for the detection, quantification, and differentiation of four Solanaceae: potato (Solanum tuberosum), tomato (Solanum lycopersicum), pepper (Capsicum annuum), and eggplant (Solanum melongena). Sequence variability was studied first using lines and cultivars (intraspecies sequence variability), then using taxa involved in gene introgressions, and finally, using taxonomically close taxa (interspecies sequence variability). This study allowed us to design four highly specific TaqMan-MGB probes. A duplex real time PCR assay was developed for simultaneous quantification of tomato and potato. For eggplant and pepper, only simplex real time PCR tests were developed. The results demonstrated the high specificity and sensitivity of the assays. We therefore conclude that β-fructosidase can be used as an endogenous reference gene for GMO analysis. |
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Bibliography: | http://dx.doi.org/10.1021/jf073313n ark:/67375/TPS-4T3JK7XT-1 Cultivars of taxa used for the first assessment of the specificity (Table 1); illustration of sequence alignment of the region targeted in the Solananceae taxa (Figure 1 for tomato, Figure 2 for potato, Figure 3 for eggplant, and Figure 4 for pepper). These information are available free of charge via the Internet at http://pubs.acs.org. This study was financially supported by the DGCCRF (Direction Générale de la Consommation, de la Concurrence et de la Répression des Fraudes) contract no. A02543, GMO convention no. 1009. istex:974A402F205D714F93687A1BBE9631CC70551F1C ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0021-8561 1520-5118 |
DOI: | 10.1021/jf073313n |