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Five pentaplex real-time PCR systems for the efficient determination of 20 genetically modified maize traits in food

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Abstract

According to the EU and Swiss food legislation, only authorised traits of transgene plants are allowed to be imported and sold to the consumer. In order to control imports of maize products from retailers, efficient and reliable methods for the detection and quantification are a prerequisite. Actually, in the EU, more than 25 gm Maize traits were authorised and are allowed to be sold. To ensure correct labelling and exclude unauthorised gm traits from sale, five multiplex real-time polymerase chain reaction systems were developed and validated. These 20 systems for the most prevalent maize gm traits allow relative multiplex quantification and/or delta–delta Ct method quantification in routine in a reliable, cost- and time-efficient way.

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References

  1. Permingeat HR, Reggiardo MI, Vallejos RH (2002) Detection and quantification of transgenes in grains by multiplex and real-time PCR. J Agric Food Chem 50(16):4431–4436

    Article  CAS  Google Scholar 

  2. James D, Schmidt AM, Wall E, Green M, Masri S (2003) Reliable detection and identification of genetically modified maize, soybean, and canola by multiplex PCR analysis. J Agric Food Chem 51(20):5829–5834

    Article  CAS  Google Scholar 

  3. Hernández M, Rodríguez-Lázaro D, Zhang D, Esteve T, Pla M, Prat S (2005) Interlaboratory transfer of a PCR multiplex method for simultaneous detection of four genetically modified maize lines: Bt11, MON810, T25, and GA21. J Agric Food Chem 53(9):3333–3337 (4)

    Article  Google Scholar 

  4. Randhawa GJ, Chhabra R, Singh M (2009) Multiplex PCR-based simultaneous amplification of selectable marker and reporter genes for the screening of genetically modified crops. J Agric Food Chem 57(12):5167–5172

    Article  CAS  Google Scholar 

  5. Randhawa GJ, Chhabra R, Singh M (2010) Decaplex and real-time PCR based detection of MON531 and MON15985 Bt cotton events. J Agric Food Chem 58(18):9875–9881

    Article  CAS  Google Scholar 

  6. Bjarte R, Drømtorp SM, Rudi K, Heir E, Holck AL (2008) Determination of eight genetically modified maize events by quantitative, multiplex PCR and fluorescence capillary gel electrophoresis. Eur Food Res Technol 227(4):1125–1137

    Article  Google Scholar 

  7. Heide BR, Heir E, Holck A (2008) Detection of eight GMO maize events by qualitative, multiplex PCR and fluorescence capillary gel electrophoresis. Eur Food Res Technol 227(2):527–535

    Article  CAS  Google Scholar 

  8. García-Cañas V, Cifuentes A (2008) Simultaneous confirmatory analysis of different transgenic maize (Zea mays) lines using multiplex polymerase chain reaction—restriction analysis and capillary gel electrophoresis with laser induced fluorescence detection. J Agric Food Chem 56(18):8280–8286

    Article  Google Scholar 

  9. Holck A, Pedersen BO, Heir E (2010) Detection of five novel GMO maize events by qualitative, multiplex PCR and fluorescence capillary gel electrophoresis. Eur Food Res Technol 231(3):475–483

    Article  CAS  Google Scholar 

  10. Zhang C, Xu W, Zhai Z, Luo Y, Yan X, Zhang N, Huang K (2011) Universal primer-multiplex-polymerase chain reaction (UP-M-PCR) and capillary electrophoresis–laser-induced fluorescence analysis for the simultaneous detection of six genetically modified maize lines. J Agric Food Chem 59(10):5188–5194

    Article  CAS  Google Scholar 

  11. Ehlert A, Moreano F, Busch U, Engel KH (2008) Development of a modular system for detection of genetically modified organisms in food based on ligation-dependent probe amplification. Eur Food Res Technol 227(3):805–812

    Article  CAS  Google Scholar 

  12. Chen L, Guo J, Wang Q, Kai G, Yang L (2011) Development of the visual loop-mediated isothermal amplification assays for seven genetically modified maize events and their application in practical samples analysis. J Agric Food Chem 59(11):5914–5918

    Article  CAS  Google Scholar 

  13. Weller SA (2000) Detection of ralstonia solanacearumstrains with a quantitative, multiplex, real-time, fluorogenic PCR (TaqMan) assay. Appl Environ Microbiol 66(7):2853–2858

    Article  CAS  Google Scholar 

  14. Wang WY, Fang TJ (2005) Development of multiplex and quantitative PCR assay to detect genetically modified roundup ready soybean in foods. J Food Drug Anal 13(2):132–138

    Google Scholar 

  15. Köppel R, van Velsen F, Felderer N, Bucher T (2012) Multiplex real-time PCR for the detection and quantification of DNA from four transgenic soy Mon 89788, A5547-127, roundup ready, A2704-12 and lectin. Eur Food Res Technol 235(1):23–28

    Article  Google Scholar 

  16. Köppel R, Bucher T, Meuwly A, Moor D (2014) Multiplex real-time PCR method for the detection and quantification of DNA from the four transgenic soy traits DP-356043-5, DP-305423-1, MON87701, BPS-CV127-9 and lectin. Eur Food Res Technol 239(2):347–355

    Article  Google Scholar 

  17. International Organization for Standardization (2005) ISO/IEC 17025—general requirements for the competence of testing and calibration laboratories. International Organization for Standardization, Geneva

    Google Scholar 

  18. European Network of GMO Laboratories (2011) Verification of real-time PCR methods for GMO testing when implementing interlaboratory validated methods—guidance document from the European network of GMO laboratories (ENGL). Joint Research Centre—Institute for Health and Consumer Protection. 1 Luxembourg, Publications Office of the European Union

  19. International Organization for Standardization (2006) IS 24276:2006. Foodstuffs—methods of analysis for the detection of genetically modified organisms and derived products—general requirements and definitions. International Organization for Standardization, Geneva

    Google Scholar 

  20. International Organization for Standardization (2005) ISO 21569:2005 foodstuffs—methods of analysis for the detection of genetically modified organisms and derived products—qualitative nucleic acid based methods. International Organization for Standardization, Geneva

    Google Scholar 

  21. International Organization for Standardization (2005) ISO 21570:2005 foodstuffs—methods of analysis for the detection of genetically modified organisms and derived products—quantitative nucleic acid based methods. International Organization for Standardization, Geneva

    Google Scholar 

  22. Definition of minimum performance requirements for analytical methods of GMO testing (2015) CRL-GMFF. http://gmo-crl.jrc.ec.europa.eu/doc/MPR%20Report%20Application%2020_10_2015.pdf. Accessed 29 Mar 2016

  23. Pfaffl MW (2001) A new mathematical model for relative quantification in realtime RT-PCR. Nucleic Acids Res 29(9):e45

    Article  CAS  Google Scholar 

  24. Köppel R, Zimmerli F (2010) Proposal for a performance factor to characterize real-time PCR systems, evaluate premixed DNA-polymerases and assign minimal performance criteria for individual quantitative PCR runs. Eur Food Res Technol 231:727–732

    Article  Google Scholar 

  25. 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 IST application in identifying unapproved genetically modified crops in Japan. J Agric Food Chem 57:26–37

    Article  CAS  Google Scholar 

  26. Rønning SB, Vaïtilingom M, Berdal KG, Holst-Jensen A (2003) Event specific real-time quantitative PCR for genetically modified Bt11 maize (Zea mays). Eur Food Res Technol 216:347–354

    Article  Google Scholar 

  27. Nielsen CR, Berdal KG, Holst-Jensen A (2004) Characterisation of the 5′ integration site and development of an event-specific real-time PCR assay for NK603 maize from a low starting copy number. Eur Food Res Technol 219:421–427

    Article  CAS  Google Scholar 

  28. Lee S-H, Yi B-Y, Kim S-J (2009) Event-specific analytical methods for biotech maize MIR 604 and DAS-59122-7. J Sci Food Agric 89(15):2616–2624

    Article  CAS  Google Scholar 

  29. Paz JL, Garca-Muniz N, Nadal A, Esteve T, Puigdomnech P, Pla M (2006) Interlaboratory transfer of a real-time polymerase chain reaction assay for quantitative detection of genetically modified maize event TC-1507. J AOAC Int 89(5):1347–1352 (6)

    Google Scholar 

  30. Kim J-H, Kim H-Y (2009) Event-specific detection methods for genetically modified maize MIR604 using real-time PCR. Food Sci Biotechnol 18(5):1118–1123

    CAS  Google Scholar 

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Acknowledgments

We thank the Official Food Control Authority of the Canton of Zurich for providing the resources for this work.

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Authors and Affiliations

  1. Official Food Control Authority of the Canton of Zurich, Zurich, Switzerland

    René Köppel, Arthika Ganeshan, Franziska van Velsen & Thomas Bucher

Authors
  1. René Köppel
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  2. Arthika Ganeshan
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  3. Franziska van Velsen
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  4. Thomas Bucher
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Correspondence to René Köppel.

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Köppel, R., Ganeshan, A., van Velsen, F. et al. Five pentaplex real-time PCR systems for the efficient determination of 20 genetically modified maize traits in food. Eur Food Res Technol 243, 215–225 (2017). https://doi.org/10.1007/s00217-016-2737-6

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  • DOI: https://doi.org/10.1007/s00217-016-2737-6

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