Transgenes monitoring in an industrial soybean processing chain by DNA-based conventional approaches and biosensors

Abstract The development of analytical methods for genetically modified organisms (GMO) screening is of great interest. In particular, since even highly processed GMO-derived food products are covered by new European legislations, a great effort has been devoted to the application of the analytical tests to these products. This work describes a polymerase chain reaction-based qualitative screening assay and a biosensor-based approach to detect transgenes in a Roundup Ready ® soybean processing line. Roundup Ready ® soybean was specifically analyzed in eight types of processed materials – seeds, crushed seeds, expander, crude flour, proteic flour, crude oil, degummed oil and lecithin – all derived from the same initial source and produced during the manufacturing process. Specific combinations of primers were used to differentiate sequences from the whole insert. The amplification of “marker” fragments with a maximum length of 500 bp was successfully achieved both in raw material (seeds) and in partially (crushed seeds, crude and proteic flours) and highly (crude and degummed oils and fluid lecithin) processed materials. Moreover, the extraction procedure was optimised and the polymerase chain reaction-electrophoresis analysis has been implemented by a biosensor-based approach.

[1]  Sabrina Gioria,et al.  Toward metrological traceability for DNA fragment ratios in GM quantification. 1. Effect of DNA extraction methods on the quantitative determination of Bt176 corn by real-time PCR. , 2007, Journal of agricultural and food chemistry.

[2]  Esther J Kok,et al.  Traceability of genetically modified organisms , 2002, Expert review of molecular diagnostics.

[3]  Carmela Martino,et al.  A comparison of DNA extraction methods for food analysis , 2007 .

[4]  J. Lüthy,et al.  Detection strategies for food authenticity and genetically modified foods , 1999 .

[5]  Farid E Ahmed,et al.  Detection of genetically modified organisms in foods. , 2002, Trends in biotechnology.

[6]  Sara Tombelli,et al.  Quartz crystal microbalance (QCM) affinity biosensor for genetically modified organisms (GMOs) detection. , 2003, Biosensors & bioelectronics.

[7]  Sara Tombelli,et al.  Combination of amplification and post-amplification strategies to improve optical DNA sensing. , 2003, Biosensors & bioelectronics.

[8]  Sara Tombelli,et al.  Detection of fragmented genomic DNA by PCR-free piezoelectric sensing using a denaturation approach. , 2005, Journal of the American Chemical Society.

[9]  Roberto Corradini,et al.  Development of a peptide nucleic acid array platform for the detection of genetically modified organisms in food. , 2005, Journal of agricultural and food chemistry.

[10]  M. Buiatti,et al.  Genome flux in tomato auto- and auxo-trophic cell clones cultured in different auxin/cytokinin equilibria. I. DNA multiplicity and methylation levels. , 1995, Genome.

[11]  Despina P Kalogianni,et al.  Nanoparticle-based DNA biosensor for visual detection of genetically modified organisms. , 2006, Biosensors & bioelectronics.

[12]  Elke Anklam,et al.  Analytical methods for detection and determination of genetically modified organisms in agricultural crops and plant-derived food products , 2002 .

[13]  S. Kang,et al.  Microchip capillary gel electrophoresis using programmed field strength gradients for the ultra-fast analysis of genetically modified organisms in soybeans. , 2005, Journal of chromatography. A.

[14]  K. Engel,et al.  Distortion of genetically modified organism quantification in processed foods: influence of particle size compositions and heat-induced DNA degradation. , 2005, Journal of agricultural and food chemistry.

[15]  Roberto Gambari,et al.  Surface plasmon resonance for detection of genetically modified organisms in the food supply. , 2006, Journal of AOAC International.

[16]  Rolf Meyer,et al.  Polymerase chain reaction (PCR) in the quality and safety assurance of food: Detection of soya in processed meat products , 1996, Zeitschrift fur Lebensmittel-Untersuchung und -Forschung.

[17]  Yves Bertheau,et al.  Trends in analytical methodology in food safety and quality: monitoring microorganisms and genetically modified organisms , 2007 .

[18]  Philipp Weller,et al.  The effect of processing parameters on DNA degradation in food , 2003 .

[19]  C. Peano,et al.  Development of a peptide nucleic acid polymerase chain reaction clamping assay for semiquantitative evaluation of genetically modified organism content in food. , 2005, Analytical biochemistry.

[20]  R. Marchelli,et al.  A microarray platform for parallel detection of five transgenic events in foods: a combined polymerase chain reaction-ligation detection reaction-universal array method. , 2005, Journal of agricultural and food chemistry.

[21]  J. Doyle,et al.  Isolation of plant DNA from fresh tissue , 1990 .

[22]  K. Engel,et al.  Quantification of DNA from genetically modified organisms in composite and processed foods , 2006 .

[23]  M. Passamano,et al.  QCM DNA-sensor for GMOs detection , 2006 .

[24]  H Schimmel,et al.  Detection and traceability of genetically modified organisms in the food production chain. , 2004, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[25]  Patrick Philipp,et al.  A Microarray-based Detection System for Genetically Modified (GM) Food Ingredients , 2006, Plant Molecular Biology.

[26]  Shuifang Zhu,et al.  Screening genetically modified organisms using multiplex-PCR coupled with oligonucleotide microarray. , 2006, Biosensors & bioelectronics.