The detection of nucleotide sequences specific for genetically modifiedorganisms (GMOs) in raw and processed food is based on different technologicalstrategies, such as the extraction of DNA and the amplificationby polymerase chain reaction (PCR), which allow to obtain qualitativeand quantitative information. We developed a multiplex PCR-basedDNA assay for simultaneously detecting multiple target sequences ingenetically modified (GM) soybean (Roundup ReadyTM). Internal controltarget (lectin gene) was included both to assess the efficiency of all reactionsand eliminating any false negatives. The post-PCR analysis wascarried out by 2.5% agarose gel electrophoresis followed by ethidiumbromide staining and densitometric analysis. The multiplex PCR method,showing high sensitivity and specificity, was tested on DNA extractedfrom certified reference samples containing GM soybean, and fromfood samples (feeds, food supplements, etc.). Comparison of this methodwith a quantitative evaluation, carried out by real-time PCR, suggests apossible utilization of the multiplex approach for semi-quantitative determinations.The method reported in this work can considerably reduce thetime and the costs of the GM soybean detection, especially in the screeningof a large number of food samples.[...]
A multiplex PCR-based assay for the detection of genetically modified soybean
DAINESE, Enrico;ANGELUCCI, Clotilde;COZZANI, Ivo
2004-01-01
Abstract
The detection of nucleotide sequences specific for genetically modifiedorganisms (GMOs) in raw and processed food is based on different technologicalstrategies, such as the extraction of DNA and the amplificationby polymerase chain reaction (PCR), which allow to obtain qualitativeand quantitative information. We developed a multiplex PCR-basedDNA assay for simultaneously detecting multiple target sequences ingenetically modified (GM) soybean (Roundup ReadyTM). Internal controltarget (lectin gene) was included both to assess the efficiency of all reactionsand eliminating any false negatives. The post-PCR analysis wascarried out by 2.5% agarose gel electrophoresis followed by ethidiumbromide staining and densitometric analysis. The multiplex PCR method,showing high sensitivity and specificity, was tested on DNA extractedfrom certified reference samples containing GM soybean, and fromfood samples (feeds, food supplements, etc.). Comparison of this methodwith a quantitative evaluation, carried out by real-time PCR, suggests apossible utilization of the multiplex approach for semi-quantitative determinations.The method reported in this work can considerably reduce thetime and the costs of the GM soybean detection, especially in the screeningof a large number of food samples.[...]I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
