Abstract
Although PCR technology has obvious limitations, the potentially high degree of sensitivity and specificity explains why it has been the first choice of most analytical laboratories interested in detection of genetically modified (GM) organisms (GMOs) and derived materials. Because the products that laboratories receive for analysis are often processed and refined, the quality and quantity of target analyte (e.g. protein or DNA) frequently challenges the sensitivity of any detection method. Among the currently available methods, PCR methods are generally accepted as the most sensitive and reliable methods for detection of GM-derived material in routine applications.
The choice of target sequence motif is the single most important factor controlling the specificity of the PCR method. The target sequence is normally a part of the modified gene construct, for example a promoter, a terminator, a gene, or a junction between two of these elements. However, the elements may originate from wildtype organisms, they may be present in more than one GMO, and their copy number may also vary from one GMO to another. They may even be combined in a similar way in more than one GMO. Thus, the choice of method should fit the purpose. Recent developments include event-specific methods, particularly useful for identification and quantification of GM content.
Thresholds for labelling are now in place in many countries including those in the European Union. The success of the labelling schemes is dependent upon the efficiency with which GM-derived material can be detected. We will present an overview of currently available PCR methods for screening and quantification of GM-derived DNA, and discuss their applicability and limitations. In addition, we will discuss some of the major challenges related to determination of the limits of detection (LOD) and quantification (LOQ), and to validation of methods.
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References
Kleppe K, Ohtsuka E, Kleppe R, Molineux I, Khorana HG (1971) J Mol Biol 56:341–361
Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988) Science 239:487–491
Institute of Reference Materials and Measurements (IRMM), European Commission, Joint Research Centre, Geel, Belgium (2002) http://www.irmm.jrc.be/. Cited 12 Sept 2002
Agriculture and Biotechnology Strategies Canada inc (2002) GMO database. http://www.agbios.com. Cited 12 Sept 2002
Wolf C, Scherzinger M, Wurz A, Pauli U, Hübner P, Lüthy J (2000) Eur Food Res Technol 210:367–372
Hemmer W (1997) BATS-report 2/97. BATS, Basel, Switzerland
Commission Regulation 49/2000 EC (2000) Off J Europ Com-11.1.2000-No L 6, P. 0013–0014
Berdal KG, Holst-Jensen A (2001) Eur Food Res Technol 213:432–438
European Committee for Normalisation (CEN), Technical Committee 275, Working group 11, internal working documents
Studer E, Rhyner C, Lüthy J, Hübner P (1998) Z Lebensm Unters Forsch 207:207–213
Hardegger M, Brodmann P, Herrmann A (1999) Eur Food Res Technol 209:83–87
Van den Eede G, Lipp M, Eyquem F, Anklam E (2000) European Commission, Joint Research Centre, IHCP. EUR 19676 EN
Rudi K, Holck A, Matforsk, Norway, personal communication
García-Cañas V, Gonzáles R, Cifuentes A (2002) J Agric Food Chem 50:1016–1021
Burns M, Shanahan D, Valdivia H, Harris N (2003) Eur Food Res Technol (in press)
Taberlet P, Gielly L, Pautou G, Bouvet J (1991) Plant Mol Biol 17:1105–1109
Zimmermann A, Liniger M, Lüthy J, Pauli U (1998) Lebensm-Wiss u Technol 31:664–667
Ehlers B, Strauch E, Goltz M, Kubsch D, Wagner H, Maidhof H, Bendiek J, Appel B, Buhk H-J (1997) Bundesgesundhbl 4:118–121
Vaïtilingom M, Pijnenburg H, Gendre F, Brignon P (1999) J Agric Food Chem 47:5261–5266
Hernandez M, Rio A, Esteve T, Prat S, Pla M. (2001) J Agric Food Chem 49:3622–3627
Meyer R, Chardonnens F, Hübner P, Lüthy J (1996) Z Lebensm Unters Forsch 203:339–344
Busch U, Mühlbauer B, Schulze M, Zagon J (1999) Deutsche Lebensmittelrundschau Heft 2:52–56
Pietsch K, Waiblinger H-U, Brodmann P, Wurz A (1997) Deutsche Lebensm Rundsch 93:35–38
Matsuoka T, Kuribara H, Takubo K, Akiyama H, Miura H, Goda Y, Kusakabe Y, Isshiki K, Toyoda M, Hino A (2002) J Agric Food Chem 50:2100–2109
Matsuoka T, Kuribara H, Akiyama H, Miura H, Goda Y, Kusakabe Y, Isshiki K, Toyoda M, Hino A (2001) J Food Hyg Soc Japan 42:24–32
Hupfer C, Hotzel H, Sachse K, Engel K-H (1998) Z Lebensm Unters Forsch 206:203–207
Wurz A, Willmund (1997) Foods produced by means of genetic engineering. In: Schrieber, GA, Bögl KW (eds) 2nd status report. BgVV-Heft 1/1997, pp 115–117
Zimmermann A, Lüthy J, Pauli U (2000) Lebensm-Wiss u Technol 33:210–216
Rønning SB, Vaïtilingom M, Berdal KG, Holst-Jensen A (2003) Eur Food Res Technol (in press)
Holck A, Vaïtilingom M, Didierjean L, Rudi K (2002) Eur Food Res Technol 214:449–454
Hernandez M, Pla M, Esteve T, Prat S, Puigdomenech P, Ferrando A (2002) Transgenic Res (in press)
Windels P, Bertrand S, Depicker A, Moens W, Van Bockstaele E, De Loose M (2003) Eur Food Res Technol (in press)
Taverniers I, Wiendels P, Van Bockstaele E, De Loose M (2001) Eur Food Res Technol 213:417–424
Terry C, Harris N (2001) Eur Food Res Technol 213:425–431
Arumuganathan K, Earle ED (1991) Plant Mol Biol Rep 9:211–215
Kay S, Van den Eede G (2001) Nature Biotechnol 19:405
Windels P, Taverniers I, Depicker A, Van Bockstaele E, De Loose M (2001) Eur Food Res Technol 213:107–112
Prokisch J, Zeleny R, Trapmann S, Le Guern L, Schimmel H, Kramer GN, Pauwels J (2001) Fresen J Anal Chem 370:935–939
Acknowledgements
This study was financially supported by the European Commissions Framework 5, Quality of Life program, within the project "Reliable, Standardised, Specific, Quantitative Detection of Genetically Modified Food" (QLK1–1999–01301), and a grant from the Research Council of Norway (NFR 136430/130).
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Holst-Jensen, A., Rønning, S.B., Løvseth, A. et al. PCR technology for screening and quantification of genetically modified organisms (GMOs). Anal Bioanal Chem 375, 985–993 (2003). https://doi.org/10.1007/s00216-003-1767-7
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DOI: https://doi.org/10.1007/s00216-003-1767-7