Vol 45(2011) N 6 p. 894-903;
D.A. Gryadunov1*, I.A. Getman2, S.N. Chizhova2, V.M. Mikhailovich1, A.S. Zasedatelev1,2
Identification of Plant-Derived Genetically Modified Organisms in Food and Feed Using a Hydrogel Oligonucleotide Microchip1Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia
2Timiryazev Insitute of Plant Physiology Russian Academy of Sciences, Moscow, 127276 Russia
Received - 2011-03-15; Accepted - 2011-04-11
A method of multiplex polymerase chain reaction (PCR) followed by hybridization on a hydrogel oligonucleotide biochip was developed for simultaneous identification of ten different transgenic elements of plant DNA in food and feed products. The biochip contained 22 immobilized oligonucleotide probes that were intended for (1) detection of plant DNA, (2) determination of plant species (soybean, maize, potato, and rice), and (3) identification of transgenic elements, including sequences of 35S CaMV, 35S FMV, rice actin gene pro- moters, nos, 35S CaMV, ocs, pea rbcS1 gene terminators, and bar, gus, and nptII marker genes. The limit of detection was 0.5% for genetically modified (GM) soybean and maize in the analyzed samples. The tests on food and feed products using the developed approach and real-time PCR showed full agreement in determination of transgenic DNA in the samples. The proposed assay can be used for selection of GM samples by screening food and feed products for subsequent quantitative determination of GM component based on the identified transgene.
genetically modified sources, transgenic elements, multiplex PCR, biochip, hybridization