Abstract
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.
Highlights
Since the mid-1990s, the traditional gene pool available to the plant breeders has been significantly widened by genetic engineering, giving rise to a large number of transgenic cultivars [1]
The chronoamperometric detection of the specific DNA fragment of P35S was based on a sandwich hybridization assay previously optimized by our group
A binary self-assembled monolayer composed of a thiolated capture probe (CP) and p-aminothiophenol (p-ATP) was built following a backfilling strategy described elsewhere [14]
Summary
Since the mid-1990s, the traditional gene pool available to the plant breeders has been significantly widened by genetic engineering, giving rise to a large number of transgenic cultivars [1]. The increasing cultivation of genetically modified crops has been matched by a strong scientific and social debate still open [2]. In this regard, the fast pace of modern life and global markets seems to be incompatible with the time required to clarify remaining uncertainties as those related to long-term effects on human and animal health, as well as on global ecology.
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