European Food Research and Technology

, Volume 215, Issue 1, pp 59–64

Real-time multiplex PCR: An accurate method for the detection and quantification of 35S-CaMV promoter in genetically modified maize-containing food

Authors

  • Michaela Höhne
    • Nestec Ltd., Nestlé Research Center, Department of Quality and Safety Assurance, Vers-chez-les-Blanc, 1000 Lausanne 26, Switzerland
  • Christelle Santisi
    • Nestec Ltd., Nestlé Research Center, Department of Quality and Safety Assurance, Vers-chez-les-Blanc, 1000 Lausanne 26, Switzerland
  • Rolf Meyer
    • Nestec Ltd., Nestlé Research Center, Department of Quality and Safety Assurance, Vers-chez-les-Blanc, 1000 Lausanne 26, Switzerland
Original Paper

DOI: 10.1007/s00217-002-0503-4

Cite this article as:
Höhne, M., Santisi, C. & Meyer, R. Eur Food Res Technol (2002) 215: 59. doi:10.1007/s00217-002-0503-4

Abstract.

A very sensitive and new real-time multiplex PCR method for the quantification of genetically modified (GM) maize crops in food materials was developed and validated for an ABI Prism 7700 Sequence Detection System. In the assay described, fluorescence-labelled TaqMan probes were chosen to detect the amplified DNA fragments during PCR. In this multiplex approach, maize-specific DNA (zein) and 35S-CaMV promoter-specific DNA fragments are amplified in the same tube. The method was tested for the detection and quantification of the four maize events that are approved in Europe and contain the 35S-CaMV promoter: Bt11, Bt176, Mon810 and T25 maize. Quantification was based on a standard curve prepared from certified maize flour reference material prepared by the Institute for Reference Materials and Measurements. Quantification within the range of the standard curve (0.05–1% GM maize) and up to 100% was possible. Repeatability of the method for each GM maize event was determined; coefficients of variations ranged from 28–40%. In addition, three internal Nestlé laboratories successfully applied this method and comparable results were obtained.

Genetically modified organism Quantification Real-time Multiplex Maize

Copyright information

© Springer-Verlag 2002