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

Dipartimento di Biologia Evoluzionistica “Leo Pardi”., Università degli Studi di Firenze, Via Romana 17-19, Firenze, Italy
Food Chemistry (Impact Factor: 3.39). 03/2009; 113(2):658-664. DOI: 10.1016/j.foodchem.2008.07.056

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.

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    • "45% of the samples showed more than one GM sequence, which indicate that the critical conditions during the elaboration of food processing were not sufficient to degrade these GM sequences. Transgenic monitoring in an industrial soybean processing chain is of great interest since even highly processed GMO-derived food products are covered by new European legislations (Bogani et al., 2009). PCR assays with combination of primers to CaMV 35S promoter and cry1A gene showed that 65% of the samples contained a GM sequence. "
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    ABSTRACT: The soybean is a typical legume which is used to elaborate several foods around the world. Ten imported soybean seed samples were collected and planted to identify possible genetic modifications. DNA was isolated from 20 days old seedlings. After that, PCR was carried out using the 35 S, RR and cry1AB/1AS primer pairs. Nine soybean samples were identified as genetically modified. These soybeans were used to prepare six different soybean foods. During food processing, critical steps were identified; such as drastic changes in temperature and pH. A sample was taken from these critical points and from the final product for DNA extraction and PCR amplification. In most of the samples the presence of the CaMV 35 S promoter and the gene cry1A were identified. In addition, presence of transgenic proteins was evaluated using ELISA-DAS assay. Presence of CP4 EPSPS proteins was detected in most of the studied soybean food samples except in yogurt and tofu. No cry1AB/1AC proteins were identified in any of the samples tested.
    Food Bioscience 04/2015; 11. DOI:10.1016/j.fbio.2015.04.002
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    • "With the increased understanding of the structure, organization, sequence and function of nucleic acid molecules, sequence-specific DNA detection has become increasingly important. Detection of specific DNA sequences is needed in many fields: the Human Genome Project is providing massive amounts of genetic information that should revolutionize our understanding and diagnosis of inherited diseases [1]; pathogens responsible for human and animal diseases, bacteria and viruses, are detectable via their unique nucleic acid sequences [2–5]; genetically modified organisms (GMOs) could also be detected via their specific nucleic acid fragments of artificial introduction [6,7]. It also holds enormous potential for the development of new and specific therapeutic procedures, new drug research and development, gene therapy, food technology, environmental sciences, etc [8–12]. "
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    ABSTRACT: As knowledge of the structure and function of nucleic acid molecules has increased, sequence-specific DNA detection has gained increased importance. DNA biosensors based on nucleic acid hybridization have been actively developed because of their specificity, speed, portability, and low cost. Recently, there has been considerable interest in using nano-materials for DNA biosensors. Because of their high surface-to-volume ratios and excellent biological compatibilities, nano-materials could be used to increase the amount of DNA immobilization; moreover, DNA bound to nano-materials can maintain its biological activity. Alternatively, signal amplification by labeling a targeted analyte with nano-materials has also been reported for DNA biosensors in many papers. This review summarizes the applications of various nano-materials for DNA biosensors during past five years. We found that nano-materials of small sizes were advantageous as substrates for DNA attachment or as labels for signal amplification; and use of two or more types of nano-materials in the biosensors could improve their overall quality and to overcome the deficiencies of the individual nano-components. Most current DNA biosensors require the use of polymerase chain reaction (PCR) in their protocols. However, further development of nano-materials with smaller size and/or with improved biological and chemical properties would substantially enhance the accuracy, selectivity and sensitivity of DNA biosensors. Thus, DNA biosensors without PCR amplification may become a reality in the foreseeable future.
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