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; Dipartimento di Chimica, Università degli Studi di Firenze, Polo Scientifico, Via della Lastruccia 3, 50019 Sesto F.No., Firenze, Italy
Food Chemistry 01/2009; 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.

1 Bookmark
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: In order to detect presence and quantity of Roundup Ready (RR) soybean in crude oil extracted from soybean seed with a different percentage of GMO seed two extraction methods were used, CTAB and DNeasy Plant Mini Kit. The amplifications of lectin gene, used to check the presence of soybean DNA, were not achieved in all CTAB extracts of DNA, while commercial kit gave satisfactory results. Comparing actual and estimated GMO content between two extraction methods, root mean square deviation for kit is 0.208 and for CTAB is 2.127, clearly demonstrated superiority of kit over CTAB extraction. The results of quantification evidently showed that if the oil samples originate from soybean seed with varying percentage of RR, it is possible to monitor the GMO content at the first stage of processing crude oil.
    Food Chemistry 02/2014; 145C:1072-1075. · 3.33 Impact Factor
  • Source
    Chapter: Soybean Oil
    03/2011: pages 59 - 105; , ISBN: 9781444339925
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: In this study, four different protocols were tested for their ability to extract DNA from blended refined vegetable oils: the in-house prepared Wizard and CTAB methods and the methods based on the use of the commercial kits Wizard® Magnetic DNA purification system for food and Nucleospin® for food. The performance of the extraction protocols was determined by end-point polymerase chain reaction (PCR) targeting the soybean lectin gene with primers suitable for the amplification of small fragments and confirmed by real-time PCR with specific hydrolysis probes. From the tested protocols, the Nucleospin method was the only one able to produce amplifiable DNA from refined vegetable oils. To verify the presence of Roundup Ready® (RR) soybean, event-specific primers were used for end-point PCR assays. The amplification of trace amounts of RR soybean by real-time PCR confirmed the label statements of two samples. The results highlight the importance of the DNA extraction protocol and the critical choice of PCR primers on processed food matrices, such as refined oils. Considering the few reports and difficulties pointed out in the literature to obtain amplifiable DNA from refined vegetable oils, the present results can be a step forward in the traceability of refined oils regarding authenticity issues and genetically modified organism detection. KeywordsSoybean oil-Vegetable oil-DNA extraction-Real-time PCR-GMO detection-Refined oil
    European Food Research and Technology 01/2010; 230(6):915-923. · 1.39 Impact Factor

Full-text (2 Sources)

Available from
May 29, 2014