Electrochemical detection of modified maize gene sequences by multiplexed labeling with osmium tetroxide bipyridine

In this report we demonstrate an approach for the electrochemical detection of four sequences from maize and genetically modified (GM) maize by means of square-wave voltammetry (SWV). After multiplexed labeling with osmium tetroxide bipyridine ([OsO4(bipy)]), the target oligonucleotides are hybridized with a complementary DNA capture probe immobilized on gold electrodes. The multiplexed labeling was performed by mixing the four target strands with the respective oligonucleotides 80% homologous to the central target recognition sequences in order to protect the latter from binding Of [OsO4(bipy)] to its thymine or cytosine residues. All components were added to the same solution. No significant decreases in SWV hybridization signals were observed after such multiplexed labeling of up to four target strands in the same reaction batch. Obtained voltammetric signals were significantly higher at 50 degrees C compared to 25 degrees C hybridization temperature and very low response was observed for non-complementary strands. Multiplexed labeling with osmium tetroxide bipyridine holds great promise for the development of simple and effective voltammetric detection protocols for GM organisms. (c) 2009 Elsevier B.V. All rights reserved.