Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
179675 | Electrochemistry Communications | 2012 | 4 Pages |
We demonstrate an approach for the electrochemical detection of genetically modified maize in real maize flour samples by means of square-wave voltammetry (SWV). After labeling the asymmetric PCR-amplified targets with osmium tetroxide bipyridine [OsO4(bipy)], they were hybridized with immobilized oligonucleotide probes on gold electrodes. We could detect the maize genes ivrp and SSIIb in near isogenic maize. The transgene cryIa/b and the MON810 specific fragment were detected in all transgenic maize samples, down to a content of 0.6% of MON810 in mixed samples. While it was possible to detect all sequences in the samples containing 100% near isogenic or respectively transgenic maize after a hybridization time of less than 10 min, a hybridization time of 30 min was necessary for the detection of the genetic modifications in samples containing only 0.9 to 0.6% of transgenic maize. No significant detection of the transgene cryIa/b or MON810 was possible when only 0.5% of transgenic maize was present in the sample, most likely due to insufficient amplification of the template DNA.
Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► Asymmetric PCR (one primer in excess) to produce single-stranded DNA for electrochemical labeling. ► Labeling of PCR-amplified ssDNA with Osmium tetroxide bipyridine. ► Detection of 0.6% of MON 810 besides the wildtype. ► Coupling of asymmetric PCR with voltammetric hybridization detection. ► Rapid analysis of genetically modified plants at the trace level is an urgent topic in crop and food sectors.