Microarray technology for multiplex detection of genetically modified organisms (GMOs) in food. In this PhD thesis we verified the feasibility of coupling multiplex PCR with LDR/universal array technology to the simultaneous detection of five different GMOs (RR soybean, MON810, Bt176, Bt11 and GA21 maize) and two endogenous genes (lectin and zein genes), after their multiplexed amplification. The results obtained in different experiments demonstrate the high molecular specificity of LDR probes and the possibility to apply this type of approach to complex samples. Although LDR/universal array approach is purely qualitative, it allows the screening for the presence of GMOs with a detection limit well below the European regulations requirements. Furthermore the successful amplification of processed foods, such as biscuits, and their detection with the LDR/universal array platform increases the robustness of the method proposed that can be considered a fast and reliable way of screening GMOs in raw material as well as in foodstuffs if combined with a proper DNA extraction method. Finally, we demonstrated that LDR/universal array platform can be considered an appropriate tool for parallel analysis of several samples increasing throughput.
Microarray technology for multiplex detection of genetically modified organisms (GMOs) in food / R. Bordoni ; Massimo Luzzana, Maria Luisa Villa. DIPARTIMENTO DI SCIENZE E TECNOLOGIE BIOMEDICHE, 2007. 19. ciclo, Anno Accademico 2005/2006.
Microarray technology for multiplex detection of genetically modified organisms (GMOs) in food
R. Bordoni
2007
Abstract
Microarray technology for multiplex detection of genetically modified organisms (GMOs) in food. In this PhD thesis we verified the feasibility of coupling multiplex PCR with LDR/universal array technology to the simultaneous detection of five different GMOs (RR soybean, MON810, Bt176, Bt11 and GA21 maize) and two endogenous genes (lectin and zein genes), after their multiplexed amplification. The results obtained in different experiments demonstrate the high molecular specificity of LDR probes and the possibility to apply this type of approach to complex samples. Although LDR/universal array approach is purely qualitative, it allows the screening for the presence of GMOs with a detection limit well below the European regulations requirements. Furthermore the successful amplification of processed foods, such as biscuits, and their detection with the LDR/universal array platform increases the robustness of the method proposed that can be considered a fast and reliable way of screening GMOs in raw material as well as in foodstuffs if combined with a proper DNA extraction method. Finally, we demonstrated that LDR/universal array platform can be considered an appropriate tool for parallel analysis of several samples increasing throughput.Pubblicazioni consigliate
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