Skip to main content

Detection of Genetically Modified Plants in Seeds, Food and Feed

  • Chapter
  • First Online:
Genetic Modification of Plants

Part of the book series: Biotechnology in Agriculture and Forestry ((AGRICULTURE,volume 64))

Abstract

Different techniques and analytical strategies are applied for detecting and quantifying the presence of genetically modified (GM) plants in food and feed products or in seeds. DNA-based detection is performed by qualitative PCR or by quantitative real-time PCR, whereas for protein-based detection immunoassays such as lateral flow devices and ELISA are applied. The testing strategy for GMO detection is constituted of a series of steps starting with a screening for frequently inserted genetic elements and gene constructs, followed by specific identification of the GM plant event and completing the analysis with the quantification of the relative amount of the GM plant event present in a given sample. This chapter outlines also the challenges currently emerging by stacked events or by the incidences of unauthorised GM plants. It provides Information on guidance documents and databases for validated detection methods used across routine control laboratories today.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Institutional subscriptions

References

  • AGBIOS (2008) http://www.agbios.com/dbase.php. Accessed 23 Jan 2009

  • Ahn S, Costa J, Emanuel JR (1996) PicoGreen quantitation of DNA: effective evaluation of samples pre- or post-PCR. Nucleic Acids Res 13:2623–2625

    Article  Google Scholar 

  • Akiyama H, Watanabe T, Wakabayashi K, Nakade S, Yasui S, Sakata K, Chiba R, Spiegelhalter F, Hino A, Maitani T (2005) Quantitative detection system for corn sample containing combined-trait genetically modified corn. Anal Chem 77:7421–7428

    Article  PubMed  CAS  Google Scholar 

  • Anklam E,Gadani F, Heinze P, Pijnenburg H, Van den Eede G (2002) Analytical methods for Detection and determination of genetically modified organisms (GMOs) in agricultural crops and plant-derived food products. Eur Food Res Technol 214:3–26

    Article  CAS  Google Scholar 

  • Arumuganathan K, Earle ED (1991) Nuclear content of some important plant species. Plant Mol Biol Rep 9:208–218

    Article  CAS  Google Scholar 

  • Bennett MD, Leitch IJ (2005) Plant DNA C-values database (rel. 4.0, Oct 2005). Royal Botanic Garden at Kew, London. http://data.kew.org/cvalues/homepage.html. Accessed 23 Jan 2009

  • Block A, Schwarz G (2003) Validation of different genomic and cloned DNA calibration standards for construct-specific quantification of LibertyLink in rapeseed by real-time PCR. Eur Food Res Technol 216:421–427

    CAS  Google Scholar 

  • Bonfini L, Moens W, Ben E, Querci M, Aygun B, Corbisier P, Morisset D, Zel J, Van den Eede G (2007) Analytes and related PCR primers used for GMO detection and quantification. JRC Sci Tech Rep 2007. EUR 23059 EN

    Google Scholar 

  • Broothaerts W, Corbisier P, Schimmel H, Trapmann S, Vincent S, Emons H (2008) A single nucleotide polymorphism (SNP839) in the adh1 reference gene affects the quantitation of genetically modified maize (Zea mays L.). J Agric Food Chem 56:8825–8831

    Article  PubMed  CAS  Google Scholar 

  • Bruderer S, Leitner KE (2003) Genetically modified (GM) crops: molecular and regulatory details, ver 2. Centre for Biosafety Assessment, Technology and Sustainability, Geneva. http://www.bats.ch/gmo-watch/GVO-report140703.pdf. Accessed 30 Jun 2003

  • Burns M, Valdivia H (2008) Modelling the limit of detection in real-time quantitative PCR. Eur Food Res Technol 226:1513–1524

    Article  CAS  Google Scholar 

  • Cankar K, Ravnikar M, Žel J, Gruden K, Toplak N (2005) Real-time polymerase chain reaction detection of cauliflower mosaic virus to complement the 35S screening assay for genetically modified organisms. J AOAC Int 88:814–822

    PubMed  CAS  Google Scholar 

  • Cankar K, Chauvensy-Ancel V, Fortabat MN, Gruden K, Kobilinsky A, Žel J, Bertheau Y (2008) Detection of nonauthorized genetically modified organisms using differential quantitative polymerase chain reaction: application to 35S in maize. Anal Biochem 376: 189–199

    Article  PubMed  CAS  Google Scholar 

  • CEN/TS (2006) Foodstuffs -- methods of analysis for the detection of genetically modified organisms and derived products -- sampling strategies. (CEN/TS 15568:2006) European Committee for Normalisation, Brussels

    Google Scholar 

  • Chaouachi M, Chupeau G, Berard A, McKhann H, Romaniuk M, Giancola S, Laval V, Bertheau Y, Brunel D (2008) A high-throughput multiplex method adapted for GMO detection. J Agric Food Chem 56:11596–11606

    Article  PubMed  CAS  Google Scholar 

  • Codex (2009) Joint FAO/WHO food standards programme -- codex committee on methods of analysis and sampling, thirtieth session (9–13 March 2009). Proposed draft guidelines on criteria for methods for the detection and identification of foods derived from biotechnology (CX/MAS 09/30/8). ftp://ftp.fao.org/codex/ccmas30/ma30_08e.pdf. Accessed 23 Jan 2009

  • CRL-GMFF (2009) European community reference laboratory for GM food & feed. Status of dossiers. http://gmo-crl.jrc.ec.europa.eu/statusofdoss.htm. Accessed 23 Jan 2009

  • Doležel J, BartoÅ¡ J,Voglmayr H, Greilhuber J (2003) Letter to the editor: nuclear DNA content and genome size of trout and human. Cytometry 51A:127–128

    Google Scholar 

  • Dong W, Yang L, Shen K, Kim B, Kleter GA, Marvin HJP, Guo R, Liang W, Zhang D (2008) GMDD: a database of GMO detection methods. BMC Bioinformatics 9:1–8

    Article  CAS  Google Scholar 

  • ENGL (2007) Explanatory document on the use of percentage of GM-DN copy numbers in relation to target taxon specific DNA copy numbers calculated in terms of haploid genomes as a general unit to express the percentages of GMOs. European Network of GMO Laboratories. http://engl.jrc.ec.europa.eu/docs/HGE-release.pdf. Accessed 23 Jan 2009

  • ENGL (2008) Definition of minimum performance requirements for analytical methods of GMO testing. European Network of GMO Laboratories. http://gmo-crl.jrc.ec.europa.eu/doc/Min_Perf_Requir_ Analyt_methods_131008.pdf. Accessed 23 Jan 2009

  • Ermolli M, Fantozzi A, Marini M, Scotti D, Balla B, Hoffmann S, Querci S, Paoletti C, Van den Eede G (2006) Food safety: screening tests used to detect and quantify GMO proteins. Accred Qual Assur 11:55–57

    Article  CAS  Google Scholar 

  • EU (2003a) Commission regulation (EC) no. 1829/2003 of the European parliament and of the council of 22 September 2003 on genetically modified food and feed. Off J L268:1–23

    Google Scholar 

  • EU (2003b) Commission regulation (EC) no. 1830/2003 of the European parliament and of the council of 22 September 2003 concerning the traceability and labelling of genetically modified organisms and the traceability of food and feed products produced from genetically modified organisms and amending Directive 2001/18/EC. Off J L268:24–28

    Google Scholar 

  • EU (2004) Commission recommendation 2004/787/EC of 4 October 2004 on technical guidance for sampling and detection of GMOs. Off J L348:18–26

    Google Scholar 

  • Fukuta S, Mizukami Y, Ishida A, Ueda J, Hasegawa M, Hayashi I, Hashimoto M, Kanbe M (2004) Real-time loop-mediated Isothermal amplification for the CaMV-35S promoter as a screening method for genetically modified organisms. Eur Food Res Technol 218:496–500

    Article  CAS  Google Scholar 

  • Grothaus GD, Bandla M, Currier T, Giroux R, Jenkins R, Lipp M, Shan G, Stave J, Pantella V (2007) Immunoassay as an analytical tool in agricultural biotechnology. J AOAC Int 89:913–928

    Google Scholar 

  • Hamels S, Leimanis S, Mazzara M, Bellocchi G, Foti N, Moens W, Remacle J, Van den Eede G (2007) Microarray method for the screening of EU approved GMOs by identification of their genetic elements. JRC Sci Tech Rep 2007. EUR 22935 EN

    Google Scholar 

  • Hernandez M, Duplan M-N, Bertier G, Vaïtilingom M, Hauser W, Freyer R, Pla M, Bertheau Y (2004) Development and comparison of four real-time polymerase chain reaction systems for specific detection and quantification of Zea mays L. Agric Food Chem 52:4632–4637

    Article  CAS  Google Scholar 

  • Hernandez M, Esteve T, Pla M (2005) Real-time polymerase chain reaction based assays for quantitative detection of barley, rice, sunflower and wheat. J Agric Food Chem 53:7003–7009

    Article  PubMed  CAS  Google Scholar 

  • Holland PM, Abramson RD, Watson R, Gelfand DH (1991) Detection of specific polymerase chain reaction product by utilizing the 5'–3' exonuclease activity of Thermus aquaticus DNA polymerase. Proc Natl Acad Sci USA 88:357–362

    Google Scholar 

  • Holst-Jensen A (2007) Sampling, detection, identification and quantification of genetically modified organisms (GMOs). In: Pico Y (2007) Food toxicants analysis. techniques, strategies and developments. Elsevier, Amsterdam, pp 231–268

    Google Scholar 

  • Holst-Jensen A, Rønning SB, Løvseth A, Berdal KG (2003) PCR technology for screening and quantification of genetically modified organisms (GMOs). Anal Bioanal Chem 375: 985–993

    PubMed  CAS  Google Scholar 

  • Hübner P, Waiblinger HU, Pietsch K, Brodmann P (2001) Validation of PCR methods for quantitation of genetically modified plants in food. J AOAC Int 84:1855–1864

    PubMed  Google Scholar 

  • ISO (1999) Cereals, pulses and milled products -- sampling of static batches. (ISO 13690) International Standardization Organisation, Geneva

    Google Scholar 

  • ISO (2002) Flowing cereals and milled cereal products -- automatic sampling by mechanical means. (ISO 6644) International Standardization Organisation, Geneva

    Google Scholar 

  • ISO (2004) Foodstuffs -- methods of analysis for the detection of genetically modified organisms and derived products -- protein based methods. (ISO 21572) International Standardization Organisation, Geneva

    Google Scholar 

  • ISO (2005a) Foodstuffs -- methods of analysis for the detection of genetically modified organisms and derived products -- qualitative nucleic acid based methods. (ISO 21569) International Standardization Organisation, Geneva

    Google Scholar 

  • ISO (2005b) Foodstuffs -- methods of analysis for the detection of genetically modified organisms and derived products -- quantitative nucleic acid based methods. (ISO 21570) International Standardization Organisation, Geneva

    Google Scholar 

  • ISO (2005c) Foodstuffs -- methods of analysis for the detection of genetically modified organisms and derived products -- nucleic acid extraction. (ISO 21571) International Standardization Organisation, Geneva

    Google Scholar 

  • ISO (2006) Foodstuffs -- methods of analysis for the detection of genetically modified organisms and derived products -- general requirements and definitions. (ISO 24276) International Standardization Organisation, Geneva

    Google Scholar 

  • JRC (2008) GMO methods database. http://biotech.jrc.it/home/ict/methodsdatabase.htm. Accessed 23 Jan 2009

  • LAG (2006) Concept for seed analysis for genetically modified plant content. Sub-committee for method development of the German national and federal länder, joint committee on genetic engineering (LAG). http://www.lag-gentechnik.de/concept for seed analysis_2006.pdf. Accessed 23 Jan 2009

  • Leimanis S, Hamels S, Nazé F, Mbongolo Mbella G, Sneyers M, Hochegger R, Broll H, Roth L, Dallmann K, Micsinai A, La Paz JL, Pla M, Brünen-Nieweler C, Papazova N, Taverniers I, Hess N, Kirschneit B, Bertheau Y, Audeon C, Laval V, Busch U, Pecoraro S, Neumann K, Rösel S, van Dijk J, Kok E, Bellocchi G, Foti N, Mazzara M, Moens W, Remacle J, Van den Eede G (2008) Validation of the performance of a GMO multiplex screening assay based on microarray detection. Eur Food Res Technol 227:1621–1632

    Article  CAS  Google Scholar 

  • Lipp M, Shillito R, Giroux R, Spiegelhalter F, Charlton S, Pinero D, Song P (2005) Polymerase chain reaction technology as an analytical tool in agricultural biotechnology. J AOAC Int 88:136–155

    PubMed  CAS  Google Scholar 

  • Ma BL, Subedi K, Evenson L, Stewart G (2005) Evaluation of detection methods for genetically modified traits in genotypes resistant to European corn borer and herbicides. J. Environ Sci Health B 40:633–644

    PubMed  CAS  Google Scholar 

  • Mäde D, Degner C, Grohmann L (2006) Detection of genetically modified rice: a construct-specific real-time PCR method based on DNA sequences from transgenic Bt rice. Eur Food Res Technol 224:271–278

    Article  Google Scholar 

  • Mano J, Shigemitsu N, Futo S, Akiyama H, Teshima R, Hino A, Furui S, Kitta K (2009) Real-time PCR array as a universal platform for the detection of genetically modified crops and its application in identifiying unapproved genetically modified crops in Japan. J Agric Food Chem 57:26–37

    Article  PubMed  CAS  Google Scholar 

  • Moreano F, Ehlert A, Busch, U, Engel KH (2006) Ligation-dependent probe amplification for the simultaneous event-specific detection and relative quantification of DNA from two genetically modified organisms. Eur Food Res Technol 222:479–485

    Article  CAS  Google Scholar 

  • Morisett D, Dobnik D, Hamels S, Žel J, Gruden K (2008) NAIMA: target amplification strategy allowing quantitative on-chip detection of GMOs. Nucleic Acids Res 36. doi:10.1093/nar/gkn524

    Google Scholar 

  • OECD (2006) OECD guidance for the designation of a unique identifier for transgenic plants. Series on the harmonisation of regulatory oversight in biotechnology, 23 (revised 2006). Organisation for Economic Cooperation and Development, London. http://appli1.oecd.org/olis/2002doc.nsf/43bb6130e5e86e5fc12569fa005d004c/fd7dd780ba22d433c125721f00598ce1/$file/jt03217233.pdf. Accessed 23 Jan 2009

    Google Scholar 

  • Papazova N, Malef A, Degrieck I, Van Bockstaele E, De Loose M (2005a) DNA extractability from the maize embryo and endosperm -- relevance to GMO assessment in seed samples. Seed Sci Technol 33:533–542

    Google Scholar 

  • Papazova N, Taverniers I, Degrieck I, Van Bockstaele E, Joost H, De Loose M (2005b) Real time polymerase chain reaction (PCR) quantification of T25 maize seeds -- influence of the genetic structure in the maize kernel on the quantitative analysis. Seed Sci Technol 34:321–331

    Google Scholar 

  • Pardigol A, Guillet S, Pöpping B (2003) A simple procedure for quantification of genetically modified organisms using hybrid amplicon standards. Eur Food Res Technol 216:412–420

    CAS  Google Scholar 

  • Prins TW, van Dijk JP, Beenen HG, Van Hoef AMA, Voorhuijzen MM, Schoen CD, Aarts HJM, Kok EJ (2008) Optimised padlock probe ligation and microarray detection of multiple (non-authorised) GMOs in a single reaction. BMC Genomics 9:584. doi:10.1186/1471-2164-9-584

    Article  PubMed  Google Scholar 

  • Querci M, Kleter G, Malingreau J-P, Broll H, Van den Eede G (2008) Scientific and technical contribution to the development of an overall health strategy in the area of GMOs. JRC Ref Rep EUR 23542 EN:1–56

    Google Scholar 

  • Reiting R, Broll H, Waiblinger H-U, Grohmann L (2007) Collaborative study of a T-nos real-time PCR method for screening of genetically modified organisms in food products. J Verbr Lebensm 2:116–121

    Article  CAS  Google Scholar 

  • Rho JK, Lee T, Jung SI, Kim TS, Park YH, Kim YM (2004) Qualitative and quantitative PCR methods for detection of three lines of genetically modified potatoes. J Agric Food Chem 52:3269–3274

    Article  PubMed  CAS  Google Scholar 

  • Roth L, Zagon J, Laube I; Holst-Jensen A; Broll H (2008) Generation of reference material by the use of multiple displacement amplification (MDA) for the detection of genetically modified organisms (GMOs). Food Anal Methods 1:181–189

    Article  Google Scholar 

  • Sambrook J, Russell DW (2001) Molecular cloning, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Taverniers I, Papazova N, Bertheau Y, De Loose M, Holst-Jensen A (2008) Gene stacking in transgenic plants: towards compliance between definitions, terminology, and detection within the EU regulatory framework. Environ Biosaf Res 7:197–218

    Article  CAS  Google Scholar 

  • Tengs T, Kristoffersen AB, Berdal KG, Thorstensen T, Butenko MA, Nesvold H, Holst-Jensen A (2007) Microarray-based method for detection of unknown genetic modifications. BMC Biotechnology 7:1–8

    Article  Google Scholar 

  • Theuns I, Windels P, De Buck S, Depicker A, Van Bockstaele E, De Loose M (2002) Identification and characterization of T-DNA inserts by T-DNA fingerprinting. Euphytica 123:75–84

    Article  CAS  Google Scholar 

  • UN (2000) Cartagena protocol on biosafety to the convention on biological diversity. www.bch.cbd.int. Accessed 23 Jan 2009

  • Van den Bulcke M, De Schrijver A, De Bernardi D, Devos Y, MbongoMbella A, Casi AL, Moens W, Sneyers M (2007) Detection of genetically modified plant products by protein strip testing: an evaluation of real-life samples. Eur Food Res Technol 225:49–57

    Article  Google Scholar 

  • Waiblinger HU, Ohmenhäuser M, Pietsch K, Ritter W, Steegmüller, J, Krech A, Horn P, Schroeder A (2005) Die Untersuchung von transgenem Rapspollen in Honigen mittels Real-time PCR. Dtsch Lebensm Rundsch 101:543–549

    CAS  Google Scholar 

  • Waiblinger HU, Ernst B, Anderson A, Pietsch K (2007) Validation and collaboration study of a P35S and T-nos duplex real-time PCR screening method to detect genetically modified organisms in food products. Eur Food Res Technol 226:1221–1228

    Article  Google Scholar 

  • Waiblinger HU, Boernsen B, Pietsch K (2008) Praktische Anwendung für die Routineanalytik. Screening-Tabelle für den Nachweis zugelassener und nicht zugelassener gentechnisch veränderter Pflanzen. Dtsch Lebenm Rundsch 104:261–264

    CAS  Google Scholar 

  • Xu J, Zhu S, Miao H, Huang W, Qiu M, Huang Y, Fu X, Li Y (2007) Event-specific detection of seven genetically modified soybean and maizes using multiplex-PCR coupled with oligonucleotide microarray. J Agric Food Chem 55:5575–5579

    Article  PubMed  CAS  Google Scholar 

  • Zeitler R, Pietsch K, Waiblinger HU (2002) Validation of real-time PCR methods for the quantification of transgenic contaminations in rape seed. Eur Food Res Technol 214:346–351

    Article  CAS  Google Scholar 

  • Žel J, Gruden K, Cankar K, Å tebih D, Blejec A (2007) Calculation of measurement uncertainty in quantitative analysis of genetically modified organisms using intermediate precision - a practical approach. J AOAC Int 90:582–586

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Lutz Grohmann .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Springer-Verlag Berlin Heidelberg

About this chapter

Cite this chapter

Grohmann, L. (2010). Detection of Genetically Modified Plants in Seeds, Food and Feed. In: Kempken, F., Jung, C. (eds) Genetic Modification of Plants. Biotechnology in Agriculture and Forestry, vol 64. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-02391-0_7

Download citation

Publish with us

Policies and ethics