Analytical and Bioanalytical Chemistry

, Volume 393, Issue 6, pp 1629–1638

Sensitive and highly specific quantitative real-time PCR and ELISA for recording a potential transfer of novel DNA and Cry1Ab protein from feed into bovine milk

  • Patrick Guertler
  • Vijay Paul
  • Christiane Albrecht
  • Heinrich H. D. Meyer
Original Paper

DOI: 10.1007/s00216-009-2667-2

Cite this article as:
Guertler, P., Paul, V., Albrecht, C. et al. Anal Bioanal Chem (2009) 393: 1629. doi:10.1007/s00216-009-2667-2

Abstract

To address food safety concerns of the public regarding the potential transfer of recombinant DNA (cry1Ab) and protein (Cry1Ab) into the milk of cows fed genetically modified maize (MON810), a highly specific and sensitive quantitative real-time PCR (qPCR) and an ELISA were developed for monitoring suspicious presence of novel DNA and Cry1Ab protein in bovine milk. The developed assays were validated according to the assay validation criteria specified in the European Commission Decision 2002/657/EC. The detection limit and detection capability of the qPCR and ELISA were 100 copies of cry1Ab μL−1 milk and 0.4 ng mL−1 Cry1Ab, respectively. Recovery rates of 84.9% (DNA) and 97% (protein) and low (<15%) imprecision revealed the reliable and accurate estimations. A specific qPCR amplification and use of a specific antibody in ELISA ascertained the high specificity of the assays. Using these assays for 90 milk samples collected from cows fed either transgenic (n = 8) or non-transgenic (n = 7) rations for 6 months, neither cry1Ab nor Cry1Ab protein were detected in any analyzed sample at the assay detection limits.

https://static-content.springer.com/image/art%3A10.1007%2Fs00216-009-2667-2/MediaObjects/216_2009_2667_Figa_HTML.gif
Figure

Schematic formats for quantitative real-time PCR and ELISA for the quantification of cry1Ab DNA and Cry1Ab protein

Keywords

Bovine milkELISAMON810Quantitative real-time PCRValidation

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Patrick Guertler
    • 1
  • Vijay Paul
    • 1
  • Christiane Albrecht
    • 1
    • 2
  • Heinrich H. D. Meyer
    • 1
  1. 1.Physiology WeihenstephanTechnische Universität MünchenFreisingGermany
  2. 2.Institute of Biochemistry and Molecular MedicineUniversity of BernBernSwitzerland