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Development of a quadruplex-real-time-PCR for screening food for genetically modified organisms

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Abstract

With the ever increasing number of genetically modified plants authorized worldwide, including in the European Union, high throughput detection methods need to be developed. In this paper, a quadruplex-real-time-PCR method is described which allows rapid and simultaneous screening of food for the presence of target DNA sequences from the cauliflower mosaic virus 35S promoter, the NOS terminator from Agrobacterium tumefaciens, the soya reference lectin gene and the maize reference alcool dehydrogenase gene (adh). Three of the four primers and probe combinations have already been published elsewhere, whereas primers and probe for NOS terminator-PCR were developed in-house. Validation data show sensitivities down to five copies for 35S promoter and NOS terminator PCR, even when target sequences of the competing PCRs are in large excess. Thorough adjustment of primer and probe concentrations allowed high individual PCR efficiencies with negligible physical cross-talk between the four detection channels. This method provides a basis for a rapid screening of food for the most frequently used regulatory elements present in GM crops authorized for food in the European Union. In addition it provides information about the presence of species which are possibly genetically modified.

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References

  1. Anklam E, Gadani F, Heinze P, Pijnenburg H, Van den Eede G (2002) Eur Food Res Technol 214:3–26

    Article  CAS  Google Scholar 

  2. Holst-Jensen A, Rønning S, Lovseth A, Berdal K (2003) Anal Bioanal Chem 375:985–993

    CAS  Google Scholar 

  3. Lipp M, Shillito R, Giroux R, Spiegelhalter F, Charlton S, Pinero D, Song P (2005) J AOAC Int 88(1):136–155

    CAS  Google Scholar 

  4. Commission regulation (EC) (2003) No. 1829/2003 OJ No. L268/1, 18.10.2003, Art24.2-3

  5. Commission regulation (EC) (2004) No. 641/2004 OJ No. L102/14, 7.4.2004

  6. Höhne M, Santisi CR, Meyer R (2002) Eur Food Res Technol 215:59–64

    Article  Google Scholar 

  7. Fernandez S, Charles-Delobel C, Geldreich A, Berthier G, Boyer F, Colonnier C, Coué-Philippe G, Diolez A, Duplan MN, Kebdani N, Romaniuk M, Feinberg M, Bertheau Y (2005) J AOAC Int 88(2):547–557

    CAS  Google Scholar 

  8. Feinberg M, Fernández S, Cassard S, Charles-Delobel C, Bertheau Y (2005) J AOAC 88(2):558–573

    CAS  Google Scholar 

  9. Reiting R, Broll H, Waiblinger HU, Grohmann L (2007) J Verbr Lebensm 2:116–121

    Article  CAS  Google Scholar 

  10. Waiblinger HU, Boernsen B, Pietsch K (2008) Deut Lebensm Rdsch 104:261–264

    CAS  Google Scholar 

  11. Xu J, Miao H, Wu H, Huang W, Tang R, Qiu M, Wen J, Zhu S, Li Y (2006) Biosens Bioelectron 22(1):71–77

    Article  CAS  Google Scholar 

  12. Xu J, Zhu S, Miao H, Huang W, Qiu M, Huang Y, Fu X, Li Y (2007) J Agric Food Chem 55:5575–5579

    Article  CAS  Google Scholar 

  13. Rudi K, Rud I, Holck A (2003) Nucleic Acid Res 31(11):e62

    Article  Google Scholar 

  14. Leimanis S, Hernández M, Fernández S, Boyer F, Burns M, Bruderer S, Glouden T, Harris N, Kaeppeli O, Philipp P, Pla M, Puigdomenech P, Vaitilingom M, Bertheau Y, Remacle J (2006) Plant Mol Biol 61:123–139

    Article  CAS  Google Scholar 

  15. Germini A, Rossi S, Zanetti A, Corradini R, Fogher C, Marchelli R (2005) J Agric Food Chem 53:3958–3962

    Article  CAS  Google Scholar 

  16. Xu X, Li Y, Zhao H, Wen S, Wang S, Huang J, Huang K, Luo Y (2005) J Agric Food Chem 53:3789–3794

    Article  CAS  Google Scholar 

  17. Schmidt A-M, Sahota R, Pope D, Lawrence S, Belton M, Rott M (2008) J Agric Food Chem 56:6791–6800

    Article  CAS  Google Scholar 

  18. Peano C, Bordoni R, Gulli M, Mezzelani A, Samson M, de Bellis G (2005) Marmiroli N Anal Biochem 346:90–100

    Article  CAS  Google Scholar 

  19. Bordoni R, Germini A, Mezzelani A, Marchelli R, De Bellis G (2005) J Agric Food Chem 53:912–918

    Article  CAS  Google Scholar 

  20. Heide B, Heir E, Holck A (2008) Eur Food Res Technol 227:527–535

    Article  CAS  Google Scholar 

  21. Heide B, Dromtorp S, Rudi K, Heir E, Holck A (2008) Eur Food Res Technol 227:1125–1137

    Article  CAS  Google Scholar 

  22. Garcia-Canas V, Cifuentes A (2008) J Agric Food Chem 56:8280–8286

    Article  CAS  Google Scholar 

  23. Permingeat HR, Reggiardo MI, Vallejos RH J (2002) Agric Food Chem 50:4431–4436

    Article  CAS  Google Scholar 

  24. Burns M, Shanahan D, Valdivia H, Harris N (2003) Eur Food Res Technol 216:428–433

    CAS  Google Scholar 

  25. James D, Schmidt A-M, Wall E, Green M, Masri S (2003) J Agric Food Chem 51:5829–5834

    Article  CAS  Google Scholar 

  26. Huang H-Y, Pan T-M (2004) J Agric Food Chem 52:3264–3268

    Article  CAS  Google Scholar 

  27. Germini A, Zanetti A, Salati C, Rossi S, Forré C, Schmid S, Marchelli R (2004) J Agric Food Chem 52:3275–3280

    Article  CAS  Google Scholar 

  28. Hernández M, Rodriguez D, Zhang D, Esteve T, Pla M, Prat S (2005) J Agric Food Chem 53:3333–3337

    Article  Google Scholar 

  29. Onishi M, Matsuoka T, Kodama T, Kashiwaba K, Futo S, Akiyama H, Maitani T, Furui S, Oguchi T, Hino A (2005) J Agric Food Chem 53:9713–9721

    Article  CAS  Google Scholar 

  30. Moreano F, Ehlert A, Busch U, Engel K-H (2006) Eur Food Res Technol 222:479–485

    Article  CAS  Google Scholar 

  31. Singh K, Ojha A, Kachru N (2007) J AOAC Int 90(6):1517–1525

    CAS  Google Scholar 

  32. Ehlert A, Moreano F, Busch U, Engel K-H (2008) Eur Food Res Technol 227:805–812

    Article  CAS  Google Scholar 

  33. Moreano F (2007) Arch Lebensmittelhyg 58:43–46

    CAS  Google Scholar 

  34. Foti N, Onori R, Donnarumma De, Santis B, Miraglia M (2006) Eur Food Res Technol 222:209–216

    Article  CAS  Google Scholar 

  35. Waiblinger H-U, Ernst B, Anderson A, Pietsch K (2008) Eur Food Res Technol 226:1221–1228

    Article  CAS  Google Scholar 

  36. Zhang H, Yang L, Guo J, Li X, Jiang L, Zhang D (2008) J Agric Food Chem 56:5514–5520

    Article  CAS  Google Scholar 

  37. Köppel Ruf, Zimmerli F, Breitenmoser A (2008) Eur Food Res Technol 227:1199–1203

    Article  Google Scholar 

  38. Berdal K, Holst-Jensen A (2001) Eur Food Res Technol 213:432–438

    Article  CAS  Google Scholar 

  39. Association française de normalisation (AFNOR), Norme expérimentale XP V03-020-2, annex D. http://www.afnor.fr

  40. European network of GMO Laboratories (2005). http://gmo-crl.jrc.ec.europa.eu/

  41. BATS Genetically modified crops: molecular and regulatory details. (http://www.bats.ch-gmo-watch-GVO-report140703.pdf.url)

  42. Shresta HK, Hwu K-K, Wang S-J, Liu L-F, Chang M-C (2008) J Agric Food Chem 56:8962–8968

    Article  Google Scholar 

  43. Leimanis S, Hamels S, Nazé F, Mbella G, Sneyers M, Hochegger R, Broll H, Roth L, Dallmann K, Micsinai A, La Paz J, 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) Eur Food Res Technol 227:1621–1632

    Article  CAS  Google Scholar 

Download references

Acknowledgment

The present work was funded by the “Fonds National de la Recherche” of the “Ministère de la Culture, de l’Education Supérieure et de la Recherche” of Luxembourg

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Authors and Affiliations

  1. Laboratoire National de Santé 1A, rue Auguste Lumière, 1950, Luxembourg, Luxembourg

    Thomas Gaudron, Claude Peters, Elodie Boland & Gilbert Moris

  2. Centre de Recherche Public de la Santé, 84 Val Fleuri, 1526, Luxembourg, Luxembourg

    André Steinmetz

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  1. Thomas Gaudron
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  2. Claude Peters
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  3. Elodie Boland
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  4. André Steinmetz
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  5. Gilbert Moris
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Correspondence to Gilbert Moris.

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Gaudron, T., Peters, C., Boland, E. et al. Development of a quadruplex-real-time-PCR for screening food for genetically modified organisms. Eur Food Res Technol 229, 295–305 (2009). https://doi.org/10.1007/s00217-009-1045-9

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  • DOI: https://doi.org/10.1007/s00217-009-1045-9

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